ABCMdb

ABCC1 p.Arg433Ser

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PMID: 14750175 [PubMed] Mizuarai S et al: "Single nucleotide polymorphisms result in impaired membrane localization and reduced atpase activity in multidrug transporter ABCG2."

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15 Homozygous CC patients expressed lower P-gp in the intestine compared to TT patients, resulting in increased digoxin plasma concentration after orally administered digoxin.21 Another report showed that a naturally occurring mutation of R433S in MRP1 caused increased organic anion transport and decreased doxorubicin resistance.22 Several groups have reported naturally occurring ABCG2 SNPs in various ethnic populations, including Caucasian, Asian and African.23-27 In those reports, polymorphisms at V12M and Q141K occurred at high frequency in most of the ethnic populations. Login to comment

PMID: 16337740 [PubMed] Cervenak J et al: "The role of the human ABCG2 multidrug transporter and its variants in cancer therapy and toxicology."

No. Sentence Comment

103 Another report showed that a naturally occurring mutation of R433S in MRP1 caused decreased organic anion transport and J. Cervenak et al. / Cancer Letters 234 (2006) 62-72 65 increased doxorubicin resistance [73]. Login to comment

PMID: 16766035 [PubMed] Cascorbi I et al: "Role of pharmacogenetics of ATP-binding cassette transporters in the pharmacokinetics of drugs."

No. Sentence Comment

852 Table 5 Frequency of ABCC1 genetic variants in different populations, position on DNA, putative effect, and frequencies (according to Le Saux et al., 2000; Ito et al., 2001; Moriya et al., 2002; Conrad et al., 2002; Oselin et al., 2003b; Wang et al., 2004) Position/ Nucleotide Aminoacid or effect Orientals Caucasians Function 128G>C C43S 0.01 - elevateda 218C>T T73I 0.00-0.04 - 257C>T S92F 0.00 0.00 decreaseda 350C>T T117M - 0.02 (decreased)a 689G>A R230N 0.00 0.00 (decreased)a 816G>A synonymous - 0.04 825T>C synonymous - 0.30 1057G>A V353M 0.00 0.005 elevateda 1299G>T R433S - 0.01 elevated Vmax of doxorubicin, decreased transport of LTC4 a,b 1684T>C synonymous - 0.80 1898G>A R633Q - 0.01 (decreased)a 2012G>T G671V - 0.03 doxorubicine-induced cardiomyopathyc 2168G>A R723Q 0.01-0.07 - decreaseda 2965G>A A989T 0.00 0.005 (decreased)a 3140G>C C1047S 0.00 0.00 3173G>A R1058Q 0.01 - 4002G>A synonymous - 0.28 4535C>T S1512L - 0.03 decreaseda a Letourneau et al. (2005). Login to comment

PMID: 17323126 [PubMed] Huang Y et al: "Pharmacogenetics/genomics of membrane transporters in cancer chemotherapy."

No. Sentence Comment

124 Moreover, Letourneau et al. examined 10 non-synonymous ABCC1 SNPs to determine Table 2 Summary of genetic variants in ABC transporters ABCB1, ABCC1, ABCC2 and ABCG2 involved in cancer chemotherapy Variants (location, effect) Phenotype Drug Sample Reference ABCB1 +103T>C (5'flanking, non-coding) Increased transcription Doxorubicin vincristine osteosarcoma Stein et al., 1994 [19] +8T>C (5'flanking, non-coding) Unknown Leukemia Rund et al., 1999 [21] 1236C>T (exon12, synonymous) Higher expression AML blasts Illmer et al., 2002 [47] Lower clearance Irinotecan Cancer patients Sai et al., 2003 [44] Higher exposure Irinotecan, SN-38 Cancer patients Mathijssen et al., 2003 [45] 2677G>T/A (exon21, A893S/T) Lower expression placenta Tanabe et al., 2001 [42] Lower expression placenta Hitzl et al., 2004 [37] Higher expression AML blasts Illmer et al., 2002 [47] Allele specific expression Cell lines, lymphoma Mickley et al., 1998 [22] Lower clearance Irinotecan Cancer patients Sai et al., 2003 [44] Survival leukemia Illmer et al., 2002 [47] Survival leukemia van den Heuvel-Eibrink et al., 2001 [48] Worse survival AML blasts Kim et al., 2006 [10] Higher efficacy Paclitaxel Ovarian cancer Green et al., 2006 [50] 2995G>A (exon24, A999T) None Cell lines, lymphoma Mickley et al., 1998 [22] 3435C>T (exon26, synonymous) Lower expression Duodenal protein Hoffmeyer et al., 2000 [26] Lower expression placenta Hitzl et al., 2004 [37] Higher expression Intestine mRNA Nakamura et al., 2002 [32] Higher expression AML blasts Illmer et al., 2002 [47] Lower clearance Irinotecan Cancer patients Sai et al., 2003 [44] Lower efflux Digoxin CD56+ NK cells Hitzl et al., 2001 [27] Higher plasma level Digoxin Healthy volunteers Hoffmeyer et al., 2000 [26] Higher AUC Cyclosporin transplant patients Bonhomme-Faivre et al., 2004 [36] Lower CNS relapse Cancer patients Stanulla et al., 2005 [46] Better survival leukemia Illmer et al., 2002 [47] Higher efficacy Breast cancer Kafka et al., 2003 [49] Higher activity, worse survival AML Kim et al., 2006 [10] Better survival Platinums Esophageal cancer Wu et al., 2006 [43] No difference Docetaxel patients Puisset et al., 2004 [41] No difference Irinotecan Cancer patients Mathijssen et al., 2004 [39] No difference Vincristine patients Plasschaert et al., 2004 [40] No difference colon Taniguchi et al., 2003 [24] ABCC1 -260G>C (5'flanking, non-coding) Higher activity Transfected cell line Wang et al., 2005 [62] Table 2 (Continued) Variants (location, effect) Phenotype Drug Sample Reference 128G>C (exon2, C43S) Reduced resistance Vincristine, arsenite Transfected cell line Leslie et al., 2003 [60] 1299G>T (exon10, R433S) Reduced transport of LTC4, increased resistance to doxorubicin Leukotriene C4, doxorubicin Transfected cell line Conrad et al., 2002 [59] 2012G>T (exon16, G671V) No change in activityLeukotriene C4 Transfected cell line Conrad et al., 2001 [58] Heart toxicity Doxorubicin nLon-Hodgkin lymphoma Wojnowski et al., 2005 [63] 2965G>A (exon22, A989T) Reduced transport Estradiol 17β-glucuronide Transfected cell line Letourneau et al., 2005 [61] ABCC2 1271A>G (exon10, R421G) Reduced drug elimination, increased nephrotoxicity Methotrexate One lymphoma patient Hulot et al., 2005 [79] 3972C>T (exon28, nonsynonymous) Reduced drug clearance Irinotecan Cancer patients Innocenti et al., 2004 [80] ABCG2 376C>T (exon4, Q126stop) Reduced transport Porphyrin Trensfected cell Tamura et al., 2006 [104] 421C>A (exon5, Q141K) Lower expression Transfected cell lines Imai et al., 2002 [94] Lower expression Transfected cell lines Kondo et al., 2004 [95] Lower expression Placenta Kobayashi et al., 2005 [98] Reduced ATPase activity Trensfected cell lines Mizuarai et al., 2004 [97] Higher plasma levels Diflomotecan patients Sparreboom et al., 2004 [100] Increased bioavailability Topotecan patients Sparreboom et al., 2005 [101] Increased bioavailability 9-Aminocamptothecin patients Zamboni et al., 2006 [81] Increased drug accumulation Imatinib Transfected cell lines Gardner et al., 2006 [96] Increased drug accumulation Topotecan Trensfected cell lines Imai et al., 2002 [94] No difference Imatinib patients Gardner et al., 2006 [96] No difference intestine Zamber et al., 2003 [99] No difference MTX Trensfected cell lines Kondo et al., 2004 [95] the effects on expression and function of this transporter in transfected HEK293T cells [61]. Login to comment

PMID: 18154452 [PubMed] Sharom FJ et al: "ABC multidrug transporters: structure, function and role in chemoresistance."

No. Sentence Comment

351 The G1299T polymorphism (exon 10) results in the change R433S in a cytoplasmic loop of MRP1, and was observed to increase doxorubicin resistance, but decrease transport of several organic anions [167]. Login to comment

PMID: 18464048 [PubMed] Gradhand U et al: "Pharmacogenomics of MRP transporters (ABCC1-5) and BCRP (ABCG2)."

No. Sentence Comment

73 In contrast, the decrease in LTC4 and estrone-3-sulfate transport due to the Arg433Ser substitution appeared to result in a lower Vmax (Conrad et al., 2002) rather than changes in Km. Login to comment
74 Interestingly, the same Arg433Ser variant was associated with increased doxorubicin resistance in transfected cells, demonstrating that a single amino acid substitution can result in markedly different substrate-dependent functional consequences. Login to comment
81 MRP1 (ABCC1) NH2 NBD NBD in out Membrane Cys43Ser Ser92Phe Thr117Met Arg230Gln Val353Met Arg633Gln Gly671Val Arg723Gln Arg433Ser Ala989Thr Cys1047Ser Val1146Ile Arg1058Gln Thr1401Met Ser1512Leu Thr73Ile COOH NBD NBD COOH NBD COOH NBD NBD Table1MRP1(ABCC1)singlenucleotidepolymorphisms.Location,allelefrequencyandfunctionaleffects. Positionin codingsequence Aminoacid exchangeLocation Allelefrequency EffectNCBIIDReferenceAfCaJpothers 128G>CCys43SerExon2--1[1]-Decreaseinvincristineresistance[2]rs41395947 Disruptedplasmamembranetraffickingin transfectedcells[2] 218C>TThr73IleExon2--1[1]3.7Chinese[3]Noinfluenceonexpressionandtransportin membranevesicles[4] rs41494447 257C>TSer92PheExon30a 0a 0a 0Chinese[3]Noinfluenceonexpressionandtransportin membranevesicles[4] 350C>TThr117MetExon3-100[5]--Noinfluenceonexpressionandtransportin membranevesicles[4] 689G>AArg230GlnExon70a 0a 0a 0Chinese[3]Noinfluenceonexpressionandtransportin membranevesicles[4] 1057G>AVal353MetExon90a 0.5a 0a -- 1299G>TArg433SerExon10-1.4[6]--Changesintransportandresistance[7] 1898G>AArg633GlnExon13-[8]--Noinfluenceonexpressionandtransportin membranevesicles[4] 2012G>TGly671ValExon16-2.8[6]--Noinfluenceonexpressionandtransportin membranevesicles[6] Associatedwithanthracycline-induced cardiotoxicity[9] 2168G>AArg723GlnExon17--7.3[1]5.6Chinese[3]Noinfluenceonexpressionandtransportin membranevesicles[4]noinfluenceonmRNA expressioninenterocytes(n=1)[10] rs4148356 2965G>AAla989ThrExon220a 0.5a 0a -Noinfluenceonexpressionandtransportin membranevesicles(non-significantreduction inE17βGtransport)[4] 323 3140G>CCys1047SerExon234.5a 0a 0a -Noinfluenceonexpressionandtransportin membranevesicles[4] rs13337489 3173G>AArg1058GlnExon23--1[1]-Noinfluenceonexpressionandtransportin membranevesicles[4] rs41410450 3436G>AVal1146IleExon24-----rs28706727 4102C>TThr1401MetExon29-----rs8057331 4535C>TSer1512LeuExon31-[5]--Noinfluenceonexpressionandtransportin membranevesicles[4] ReferencewithoutfrequencymeansthatSNPwasdetectedbutnofrequencydetermined. Login to comment

PMID: 19949922 [PubMed] Cascorbi I et al: "Pharmacogenetics of ATP-binding cassette transporters and clinical implications."

No. Sentence Comment

155 ABCC2 (Multidrug Resistance-Associated Protein 2) Table 6.5 Frequency of ABCC1 genetic variants in different populations, position on DNA, putative effect, and frequencies (according to (33, 77-80, 136)) Position Amino acid or effect Orientals Caucasians Function c.128G>C C43S 0.01 - Elevateda c. 218C>T T73I 0.00-0.04 - c. 257C>T S92F 0.00 0.00 Decreaseda c. 350C>T T117M - 0.02 (Decreased)a c. 689G>A R230N 0.00 0.00 (Decreased)a c. 816G>A Synonymous - 0.04 c. 825T>C Synonymous - 0.30 c. 1057G>A V353M 0.00 0.005 Elevateda c. 1299G>T R433S - 0.01 Elevated vmax of doxorubicin, decreased transport of LTC4 a,b c. 1684T>C Synonymous - 0.80 c. 1898G>A R633Q - 0.01 (Decreased)a c. 2012G>T G671V - 0.03 Doxorubicine-induced cardiomyopathyc c. 2168G>A R723Q 0.01-0.07 - Decreaseda c. 2965G>A A989T 0.00 0.005 (Decreased)a c. 3140G>C C1047S 0.00 0.00 c. 3173G>A R1058Q 0.01 - c. 4002G>A Synonymous - 0.28 c. 4535C>T S1512L - 0.03 Decreaseda References: a [81], b [77], c [84] an inducible expression of ABCC2, which contributes also to the phenomenon of drug resistance. Login to comment

PMID: 19950006 [PubMed] Sissung TM et al: "Pharmacogenetics of membrane transporters: an update on current approaches."

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67 Those studied include C43S, T73I, S92F, T117M, R230Q, V353M, R433S, R633Q, G671V, R723Q, A989T, C1047S, R1058Q, A1337T, and S1512L. Login to comment
69 However, it has been noted that C43S, R433S, and A989T result in decreased ABCB1 function [43]. Login to comment

PMID: 11721885 [PubMed] Conrad S et al: "Identification of human multidrug resistance protein 1 (MRP1) mutations and characterization of a G671V substitution."

No. Sentence Comment

78 Mutations in the MRP1 gene and median mRNA expression levels of samples with different MRP1 genotypes Position Location SNPs Change FRE EXPa Intron 7 809ϩ54 C/A Intronic 2.8 101 Exon 8b 825 T/C Silent 2.8 51 Intron 8 1040ϩ13 T/C Intronic 16.7 95 Exon 10 1299 G/T Arg433Ser 1.4 156 Intron 11b 1474-48 C/T Intronic 2.8 146 Intron 11 1474-8 T/C Intronic 9.7 72 Intron 12 1678-9 DEL T Intronic 11.1 106 Exon 13b 1684 T/C Silent 8.3 106 Exon 13 1704 C/T Silent 2.8 59 Exon 16 2012 G/T Gly671Val 2.8 43 Intron 20 2736-36 T/C Intronic 2.8 132 Intron 20 2736-18 CC/TT Intronic 2.8 132 Intron 20 2736-6 T/C Intronic 2.8 132 Intron 21 2871ϩ17 G/A Intronic 1.4 132 Intron 26 3819ϩ7 G/A Intronic 1.4 124 Exon 28b 4002 G/A Silent 2.8 95 Intron 30 4487ϩ5 A/T Intronic 2.8 137 Intron 30 4487ϩ18 A/G Intronic 1.4 142 Intron 30 4487ϩ28 C/G Intronic 2.8 137 Densitometrical analysis was performed with TINA software (Raytest, Straubenhardt, Germany), average relative MRP1 expression was set as 100% FRE, allelic frequency in %; EXP, expression level; SNP, single-nucleotide polymorphism a The average expression level of all samples was 100 Ϯ 55 b Recently described by Ito et al. 2001b Fig. 2. Login to comment
118 They are located in the second transmembrane spanning domain (R433S) and in the vicinity of the first ATP-binding site (G671V). Login to comment

PMID: 12042670 [PubMed] Conrad S et al: "A naturally occurring mutation in MRP1 results in a selective decrease in organic anion transport and in increased doxorubicin resistance."

No. Sentence Comment

2 In this study, we characterized a low-frequency (<1%) naturally occurring mutation in MRP1 expected to cause the substitution of a conserved arginine with serine at position 433 in a predicted cytoplasmic loop of the protein. Login to comment
5 In contrast, 17â- oestradiol-17â-(D-glucuronide) transport by the Arg433 Ser mutant MRP1 was similar to that by wild-type MRP1. Login to comment
7 In contrast to the reduced LTC4 and oestrone sulphate transport, stably transfected HeLa cells expressing Arg433 Ser mutant MRP1 were 2.1-fold more resistant to doxorubicin than cells expressing wild-type MRP1, while resistance to VP-16 and vincristine was unchanged. Login to comment
31 In addition, we have recreated the Arg433 Ser substitution in a MRP1 mammalian expression vector and analysed its transport and drug resistance properties after transfection into human cells. Login to comment
38 Site-directed mutagenesis The mutation Arg433 Ser was generated using the following sense primer 59P-GTGGACGCTCAGTC GTTCATGGACTTGGCC-39 (substituted nucleotides are underlined) with the USE Mutagenesis Kit (Amersham Pharmacia Biotech, PQ, Canada), according to the manufacturer`s instructions. Login to comment
79 Results A heterozygous Arg433 Ser substitution in MRP1 does not result in any overt clinical symptoms A mutation in exon 10 of the human MRP1 gene expected to result in substitution of Arg433 to Ser was found in a single individual among 91 healthy Caucasian volunteers. Login to comment
81 An Arg433 Ser substitution causes a reduction in MRP1-mediated LTC4 and oestrone sulphate transport activity, but leaves E217âG transport activity intact To determine whether MRP1-mediated transport of [3 H]LTC4, [3 H]oestrone sulphate and [3 H]E217âG was affected by a Arg433 Ser substitution, this mutation was generated in a mammalian MRP1 expression vector and transiently transfected into HEKSV293T cells. Login to comment
82 Stably transfected clonal HeLa cell lines expressing the Arg433 Ser MRP1 mutant were also generated in order to confirm the results obtained with the transient transfectants and to be able to test drug resistance in longer term assays. Login to comment
85 In contrast, levels of [3 H]E217âG uptake by the Arg433 Ser mutant MRP1 and wild-type MRP1 were similar and proportional to their relative protein expression levels (Fig. 2). Login to comment
87 Arg433 Ser mutant MRP1 is correctly routed to the plasma membrane To determine whether the decrease in LTC4 and oestrone sulphate transport caused by the Arg433 Ser substitution might be associated with a change in the trafficking of MRP1 to the plasma membrane, the subcellular localization of the mutant and wild-type MRP1 was determined by indirect immunofluorescent confocal microscopy. Login to comment
88 As shown in Fig. 4, both the wild-type and mutant proteins expressed in HeLa cells showed a pattern of strong plasma membrane staining, indicating that MRP1 trafficking in the transfected cells was unaffected by the Arg433 Ser substitution. Login to comment
89 Kinetic analysis of ATP-dependent [3 H]LTC4 and [3 H]oestrone sulphate transport by Arg433 Ser mutant MRP1 Since the initial time courses of LTC4 and oestrone sulphate uptake indicated a two-fold reduction in transport activity of the Arg433 Ser mutant MRP1, the effect of this mutation on the transport kinetics of these substrates was examined. Login to comment
91 A non-linear regression analysis of the data obtained showed small but not significant differences in the apparent Km values of the wild-type and Arg433 Ser mutant proteins for LTC4 and oestrone sulphate. Login to comment
92 In contrast, the apparent Vmax values of the Arg433 Ser mutant for both substrates were approximately two-fold lower than for wild-type MRP1 when normalized for relative levels of protein expression. Login to comment
93 Drug resistance profile of cells expressing Arg433 Ser MRP1 In addition to its ability to transport conjugated organic anions, MRP1 can also confer resistance to a variety of anticancer drugs by reducing intracellular drug accumulation [21,22]. Login to comment
94 To determine whether the Arg433 Ser substitution affected the drug resistance properties of MRP1, the drug resistance profile of Arg433 Ser MRP1 Fig. 1 Expression and ATP-dependent [3 H]LTC4 transport activity of wild-type and mutant Arg433 .Ser MRP1 in membrane vesicles prepared from transfected cells. Login to comment
95 (a) MRP1 expression levels in membrane vesicles of human embryonic kidney cells transfected with pcDNA3.1(-) as control, pcDNA3.1(-)wt-MRP1K (wtMRP1) and pcDNA3.1(-) MRP1K-Arg433 Ser (R433S) cDNA expression vectors were determined by dot blot analysis with MAb QCRL-1. Login to comment
97 (b) Time course of [3 H]LTC4 uptake in membrane vesicles shown in (a) prepared from HEKSV293T cells transfected with wild-type MRP1 (wtMRP1, j), mutant (R433S, m) and control (pcDNA3.1(-), h) cDNA expression vectors. Results shown are means (Æ SD) of triplicate determinations in a single experiment. Similar results were obtained in two additional independent experiments with vesicles from transfected HEKSV293T cells and in two additional experiments with vesicles from transfected HeLa cells. Login to comment
99 The Arg433 Ser mutation increased resistance to the cationic anthracycline agent, doxorubicin by approximately two-fold (P , 0.01) relative to wild-type MRP1 (Fig. 6a). Login to comment
100 In contrast, the sensitivity of the Arg433 Ser mutant to the Vinca alkaloid, vincristine, and the electroneutral epipodophyllotoxin, VP-16 (Fig. 6b,c) was comparable to that conferred by wild-type MRP1. Login to comment
108 Thus, it seems likely that this transport protein contains more than one Fig. 2 Expression and ATP-dependent [3 H]E217âG transport activity of wild-type and mutant Arg433 Ser MRP1 in membrane vesicles prepared from transfected cells. Login to comment
110 (b) Time course of [3 H]E217âG uptake in membrane vesicles shown in (a) (wtMRP1, j; mutant R433S, m; control pcDNA3.1(-), h). Login to comment
114 (b) Time course of [3 H]oestrone sulphate uptake in the presence of 1 mmol GSH in the membrane vesicles shown in (a) prepared from cells transfected with wild-type MRP1 (wtMRP1, j), mutant (R433S, m) and empty vector control (pcDNA3.1(-), h) cDNA expression vectors. Results shown are means (Æ SD) of triplicate determinations in a single experiment. Similar results were obtained in three additional independent experiments with vesicles from transfected HeLa cells and in two experiments with vesicles from transfected HEKSV293T cells. Login to comment
118 Of all the mutations detected, two are predicted to cause amino acid substitutions in the protein, Gly671 Val and Arg433 Ser. Login to comment
121 According to most topological models of MRP1, the second mutation, an Arg433 Ser substitution, is located close to the COOH-proximal end of the cytoplasmic loop connecting the predicted transmembrane (TM)7 to TM8 in the second membrane spanning domain (MSD2) of MRP1 [41-43]. Login to comment
123 When the properties of a recombinant Arg433 Ser MRP1 mutant expressed in transfected human cells were examined, a two-fold reduction in LTC4 and GSH-stimulated oestrone sulphate transport activity was observed even though the expression levels and membrane localization of the mutant MRP1 and wild-type MRP1 were comparable. Login to comment
125 Thus, the Vmax values for LTC4 and GSH-stimulated oestrone sulphate transport by the Arg433 Ser mutant MRP1 were approximately two-fold lower than Fig. 4 Confocal microscopy of transfected HeLa cells expressing wild-type and Arg433 Ser mutant MRP1 proteins. Login to comment
130 (b) Transfected HeLa cells expressing Arg433 Ser mutant MRP1. Login to comment
132 (a) ATP-dependent uptake of [3 H]LTC4 by the membrane vesicles shown in Fig. 1(a) was measured at various LTC4 concentrations (0.01-1 ìM) for 45 s at 23 8C [wild-type MRP1 (wtMRP1, j; and Arg433 Ser mutant MRP1 (R433S, m)]. Login to comment
135 (b) ATP-dependent uptake of [3 H]oestrone sulphate by the membrane vesicles shown in Fig. 3(a) [wild-type MRP1 (wtMRP1, j) and Arg433 Ser mutant MRP1 (R433S, m)]. Login to comment
141 In contrast, the levels of E217âG uptake by the Arg433 Ser mutant MRP1 were unchanged. Login to comment
155 HeLa cells expressing the Arg433 Ser mutant protein were observed to be two-fold more resistant to the anthracycline doxorubicin in addition to displaying a two-fold reduction in LTC4 and GSH-stimulated oestrone sulphate transport activity. Enhanced resistance was specific for this anthracycline as the resistance of cells expressing the Arg433 Ser mutant to the Vinca alkaloid, vincristine, and to the epipodophyllotoxin, VP-16, remained comparable to cells expressing wild-type MRP1. Login to comment
160 Furthermore, TM segments corresponding to those identified in P-glycoprotein as being important for substrate binding and/or transport have also been Table 1 Kinetic parameters of [3 H]LTC4 and GSH-stimulated [3 H]oestrone sulphate uptake by membrane vesicles from cells transfected with expression vectors encoding wild-type and Arg433 Ser mutant MRP1 Km (ìM) Vmax (pmol/mg/min) Normalized Vmax (pmol/mg/min) Substrate wtMRP1 R433S wtMRP1 R433S wtMRP1 R433S LTC4 0.18 Æ 0.04 0.14 Æ 0.06 121 Æ 8 71 Æ 10 121 Æ 8 59 Æ 8 Oestrone sulphate 0.89 Æ 0.07 1.10 Æ 0.34 72 Æ 2 41 Æ 4 72 Æ 2 34 Æ 3 Apparent Km and Vmax values for LTC4 and GSH-stimulated oestrone sulphate uptake were determined as described in Materials and Methods and the legend to Fig. 5. Login to comment
169 Thus, our data, demonstrating that the doxorubicin resistance of cells expressing the Arg433 Ser mutation is increased while vincristine and VP-16 resistance are unaffected, indicate that amino acid changes in this cytoplasmic loop of MRP1, similar to the comparable loop in P-glycoprotein, can selectively affect the drug resistance profile conferred by this transporter. Login to comment
171 Cells stably transfected with wild-type MRP1 (wtMRP1, j), mutant (R433S, m) and control (pcDNA3.1(-), h) cDNA expression vectors were exposed to (a) doxorubicin, (b) vincristine and (c) etoposide at the concentrations indicated for 72 h at 37 8C. Login to comment
174 100 75 50 25 0.1 1 10 100 Vincristine (nM) %Controlabsorbance 100 75 50 25 1 10 100 Etoposide (µM) %Controlabsorbance Table 2 Relative drug resistance of HeLa cells expressing wild-type and Arg433 Ser mutant MRP1 Relative resistancea Drug wtMRP1 Arg433 Ser MRP1 Ratio Doxorubicin 2.5 Æ 0.6 (n ¼ 5) 5.3 Æ 1.0 (n ¼ 8)b 2.1 Vincristine 7.0 Æ 4.1 (n ¼ 5) 10.3 Æ 5.0 (n ¼ 10) 1.5 VP-16 3.4 Æ 1.4 (n ¼ 6) 3.8 Æ 1.6 (n ¼ 6) 1.1 a The resistance of stably transfected HeLa cells was determined using a tetrazolium-based microtitre plate assay. The relative resistance factors were obtained by dividing the IC50 values for wild-type or Arg433 Ser mutant MRP1 transfected cells by the IC50 values for empty vector control transfected cells and are normalized for differences in protein expression levels as appropriate. Login to comment
177 Thus, the Arg433 Ser substitution results in an additional potential phosphorylation site which, if utilized, might in some direct or indirect way influence the activity of the protein. Login to comment
179 The MRP1 mutation predicted to cause the Arg433 Ser substitution described here was present in a cohort of 91 healthy Caucasian volunteers with an estimated allelic frequency of ,1%. Login to comment
181 However, the mutation was heterozygous in the case found and the alterations in Arg433 Ser MRP1 transport activity observed in the current study suggest that homozygous carriers should be examined before ruling out possible physiological consequences associated with this mutation in vivo. Login to comment
182 Thus, the two-fold decrease in LTC4 and GSH-stimulated oestrone sulphate transport by the Arg433 Ser mutant MRP1 in vitro raises the possibility that individuals bearing this polymorphism might be subject to a variety of biological effects, such as an impaired response to an inflammatory stimulus [30-32]. Login to comment

PMID: 12186871 [PubMed] Haimeur A et al: "Charged amino acids in the sixth transmembrane helix of multidrug resistance protein 1 (MRP1/ABCC1) are critical determinants of transport activity."

No. Sentence Comment

238 We previously showed that the naturally occurring mutation of MRP1-Arg433 to Ser predicted to be located in the fourth cytoplasmic loop in close proximity to the membrane interface of TM8 results in a 2-fold decrease in LTC4 and estrone 3-sulfate transport efficiency. Login to comment

PMID: 14965249 [PubMed] Haimeur A et al: "The MRP-related and BCRP/ABCG2 multidrug resistance proteins: biology, substrate specificity and regulation."

No. Sentence Comment

285 Thus, substitution of Arg433 with Ser in an ICL predicted to be close to TM8 of MRP1 caused by the low frequency (<1%) G1299T MRP1 polymorphism in exon 10 results in a substrate selective alteration in organic anion transport activity and drug resistance when the mutant protein is expressed in stably transfected HeLa cells [190]. Login to comment
286 E217βG transport was unaffected by the Arg433Ser mutation A fourth mutant phenotype is that where mutants exhibit a substrate-selective loss or gain of transport activity. Login to comment
290 In contrast, cells expressing the mutant MRP1-Arg433Ser were 2.5-fold more resistant to doxorubicin while resistance to VP-16 and vincristine remained comparable to cells expressing wild-type MRP1. Login to comment

PMID: 16006996 [PubMed] Conseil G et al: "Polymorphisms of MRP1 (ABCC1) and related ATP-dependent drug transporters."

No. Sentence Comment

148 Fig. 3 Exon 1 2 3 MSDMSD NBD1 MSD NBD2 C4535T(S1512L) G3173A (R1058Q) G3140C (C1047S) G2965A (A989T) G2168A (R723Q) G2012T(G671V) G1898A (R633Q) G1299T(R433S) G1057A (V353M) G689A (R230Q) C350T(T117M) C257T(S92F) C218T(T73I) C128C (C43S) (TM1-5) (TM6-11) (TM12-17) 4 5 6 7 8 9101112 1314 151617 1819 20 21 22 23 242526272829 30 31 Location of non-synonymous SNPs in the coding regions of the genes in the MRP1/ABCC1 gene. Login to comment

PMID: 16041239 [PubMed] Colombo S et al: "Influence of ABCB1, ABCC1, ABCC2, and ABCG2 haplotypes on the cellular exposure of nelfinavir in vivo."

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70 2650C > T exon 21 synonymous (p.L884L) Kim et al., 2001 md-v-110 rs9282563 c.2677G > T exon 21 p.A893S Kim et al., 2001 md-v-031 rs2032582 c.2677G > A exon 21 p.A893T Kim et al., 2001 md-v-109 IVS 21 + 14 - 17 delAATA intron 21 Epidauros md-v-092 IVS 21 + 49 T > C intron 21 Epidauros md-v-042 rs2032583 IVS 21 + 66 T > C intron 21 Epidauros md-v-108 IVS 26 - 156 T > C intron 25 Epidauros md-v-095 IVS 26 - 68 A > G intron 25 Epidauros md-v-164 c.3320A > C exon 26 p.Q1107P Cascorbi et al., 2002 md-v-033 c.3322T > C exon 26 p.W1108R Kroetz et al., 2003 md-v-225 c.3325C > T exon 26 p.L1109F Epidauros md-v-165 c 3364C > T exon 26 synonymous (p.A1132A) Hoffmeyer et al., 2000 md-v-034 c.3321T > A exon 26 p.S1141T Kim et al., 2001 md-v-035 c.3435C > T (Tag8) exon 26 synonymous (p.I1145I) Hoffmeyer et al., 2000 md-v-036 rs1045642 IVS 26 + 59 T > G intron 26 Epidauros md-v-097 rs2235047 IVS 26 + 80 T > C intron 26 Epidauros md-v-040 rs2235048 IVS 26 + 123_24 insCATG intron 26 Epidauros md-v-096 Tag 11 intron 27 Soranzo et al., 2004 rs1186746 Tag 12 intron 27 Soranzo et al., 2004 rs1186745 MRP1 (ABCC1) c.816G > A exon 8 synonymous (p.P272P) Epidauros mr-v-014 c.825T > C exon 8 synonymous (p.V275V) Saito et al., 2002 mr-v-015 rs246221 c.1062T > C exon 9 synonymous (p.N354N) Saito et al., 2002 mr-v-016 rs35587 c.1068G > A exon 9 synonymous (p.T356T) Epidauros mr-v-057 rs8187852 IVS 9 + 8 A > G intron 9 Saito et al., 2002 mr-v-017 rs35588 c.1303G > T exon 10 p.R433S Conrad et al., 2002 mr-v-018 IVS 10 + 64 C > T intron 10 Epidauros mr-v-019 MRP2 (ABCC2) g. Login to comment

PMID: 16041243 [PubMed] Letourneau IJ et al: "Functional characterization of non-synonymous single nucleotide polymorphisms in the gene encoding human multidrug resistance protein 1 (MRP1/ABCC1)."

No. Sentence Comment

7 When the MRP1/ABCC1 non-synonymous SNPs were evaluated by the SIFT algorithm using subsets of homologs and orthologs of MRP1/ABCC1, Arg230Gln, Val353Met, Arg433Ser, Gly671Val and Arg1058 mutations were predicted to be deleterious, whereas the PolyPhen algorithm predicted Ser92Phe and Gly671Val to be potentially damaging. Login to comment
28 Of these mutations, the Fig. 1 128G >C (C43S) 128G >T(T73I) 689G >A (R230Q)1057G >A (V353M) 1299G >T(R433S) 1898G >A (R633Q) 2012G >T(G671V) 2168G >A (R723Q) 3173G >A (R1058Q) 4535C >T(S1512L) 3140G >C (C1047S) 2965G >A (A989T) 350C >T(T117M) 257C >T(S92F) 313029282726252423222120181716151413121110987654321 19 MSD1 MSD1 MSD2 MSD3 MSD2 NBD1 MSD3 NBD2 TM 1 2 3 4 5 6 7 8 Val353Met Ala989Thr Cys1047Ser Arg1058Gln NBD2NBD1 Ser1512Leu Arg633Gln Arg433Ser Arg723Gln Thr73lle Thr117Met Arg230Gln Cys43Ser Ser92Phe Gly671Val 9 10 11 12 13 14 15 16 17 (a) (b) Location of reported non-synonymous single nucleotide polymorphisms (SNPs) in MRP1/ABCC1. Login to comment
35 A second mutation, Arg433Ser (1299G > T), located in cytoplasmic loop 4 (CL4) proximal to transmembrane (TM) helix 8 in the second MSD, decreased both LTC4 and estrone-3-sulfate (E13SO4) transport but increased resistance to doxorubicin [19]. Login to comment
46 The template for generating Table 1 Frequencies of non-synonymous single nucleotide polymorphisms in MRP1/ABCC1 Variant Amino acid substitution Allelic frequency Population References 128G > C Cys43Ser 0% (0/26) Japanese [16] 1% (1/96) Japanese [17] 218C > T Thr73Ile 0% (0/26) Japanese [16] 1% (1/96) Japanese [17] 3.7% (2/54) Chinese [37] 257C > T Ser92Phe 0% (0/220) Caucasian www.pharmGKB.org 0.5% (1/200) African-American 0% (0/60) Japanese 0% (0/14) Pacific-Islander 350C > T Thr117Met 1.6% (1/64) Caucasian [28] 689G > A Arg230Gln 0% (0/220) Caucasian www.pharmGKB.org 0.5% (1/200) African-American 0% (0/60) Japanese 0% (0/14) Pacific-Islander 1057G > A Val353Met 0.5% (1/220) Caucasian www.pharmGKB.org 0% (0/200) African-American 0% (0/60) Japanese 0% (0/14) Pacific-Islander 1299G > T Arg433Ser 1.4% (1/72) Caucasian [20] 0% (0/110) Caucasian [19] 1898G > A Arg633Gln 0.8% (2/234) Caucasian [29] 2012G > T Gly671Val 2.8% (2/72) Caucasian [20] 2.6% (6/234) Caucasian [29] 2168G > A Arg723Gln 3.8% (1/26) Japanese [16] 1% (1/96) Japanese [30] 7.3% (7/96) Japanese [17] 5.6% (3/54) Chinese [37] 2965G > A Ala989Thr 0.5% (1/220) Caucasian www.pharmGKB.org 0% (0/200) African-American 0% (0/60) Japanese 0% (0/14) Pacific-Islander 3140G > C Cys1047Ser 0% (0/220) Caucasian www.pharmGKB.org 4.5% (9/200) African-American 0% (0/60) Japanese 0% (0/14) Pacific-Islander 3173G > A Arg1058Gln 0% (0./26) Japanese [16] 1% (1/96) Japanese [17] 4535C > T Ser1512Leu 3.1% (2/24) Caucasian [28] Characterization of MRP1/ABCC1 variants in vitro Le´tourneau et al. 649 the Arg633Gln and Arg723Gln mutants was created by subcloning a HindIII fragment (1329 bp) encoding amino acids 517-959 into pGEM-3z [20]. Login to comment
94 Results from previously characterized non-synonymous SNPs, Cys43Ser, Arg433Ser and Gly671Val, were included where available for comparison [18-20]. Login to comment
123 Previous mutagenesis and inhibition studies have Fig. 3 Cys43Ser Thr73lle Ser92Phe Thr117Met Arg230Gln Arg433Ser Arg633Gln Gly671Val Arg723Gln Ala989Thr Cys1047Ser Arg1058Gln Ser1512Leu Cys43Ser Thr73lle Ser92Phe Thr117Met Arg230Gln Arg433Ser Arg633Gln Gly671Val Arg723Gln Ala989Thr Cys1047Ser Arg1058Gln Ser1512Leu Thr73lle Ser92Phe Thr117Met Arg230Gln Arg633Gln Arg723Gln Ala989Thr Cys1047Ser Arg1058Gln Ser1512Leu LTC4 % WT-MRP1 uptake 0 25 50 75 100 125 E217βG % WT-MRP1 uptake 0 25 50 75 100 125 150 MTX % WT-MRP1 uptake 0 25 50 75 100 125 (b) (c) (a) ATP-dependent vesicular transport of organic anions by mutant MRP1 proteins. Login to comment
128 Hatched bars represent previously published data on non-synonymous SNPs, Cys43Ser, Arg433Ser and Gly671Val, using membrane vesicles from stably transfected HeLa cells and are included for comparison [18-20]. Login to comment
134 Thus, when the subset included the human homologs MRP2, MRP3 and MRP6, the SIFT analysis predicted that the Arg433Ser, Gly671Val and Arg1058Gln substitutions would be deleterious for MRP1 function. Login to comment
135 However, when the analysis was expanded to include the six known mammalian orthologs of MRP1, the Arg230Gln, Val353Met, Arg433Ser and Gly671Val mutations were also predicted to be deleterious. Login to comment
136 The Val353Met mutation has not yet been tested in vitro; however, of the remaining five, only Arg433Ser has been found to adversely affect MRP1 function [19]. Login to comment
158 This observation may be construed as being Table 2 Conservation of the amino acids substituted by non-synonymous SNP of human MRP1/ABCC1a Protein Speciesb C43S T73I S92F T117Mc R230Q V353M R433S R633Qc G671V R723Q A989T C1047S R1058Q S1512L MRP1 Human C T S T R V R R G R A C R S Monkey C T S M R V R R G Q A C R S Dog C T S M R V R R G R A R R S Cow C A S M Q V R R G R A R R S Rat C A S M Q V R W G R A R R S Mouse C T S M H V R R G R A R R S MRP2 Human L A V T K A K R G K A I R E Monkey L A V T K A K R G K A I R E Dog L A V T K A K R G K A I Q Q Rat L A A T K V K R G K A A R E Mouse L A A T K V K V G K A T R E Rabbit L A V T K V K R G K A I R E MRP3 Human C L S M Y I R K G Q A V R A Rat C L S M L L R K G Q A L R V MRP4 Human - - - - I F K R G R Y T K Y MRP5 Human - - - - V T R S G R T R R S MRP6 Human P A A M R I R S G V A L R A CFTR Human - - - - R Y K A G K L I Q Q SUR1 Human V L L A T V Q R G E L R L E SUR2 Human V L H T Q V Q R G E I N L P Pgp Human - - - - - E K S G A G R R Q YCF1 Saccharomyces cervisiae A I L V T V K L G K S Y R G Mrp1 Caenorhabditis elegans T L D F L I R T G R G L R K Mrp2 Caenorhabditis elegans T F D I L I K T G R G I R K AtMRP2 Arabidopsis thaliana Q L R W L M S P G R R K R E AtMRP1 Arabidopsis thaliana H T A V L M S P G R R K R E a Aligned using Clustal W (http://pbil.univ-lyon1.fr/). Login to comment

PMID: 16230346 [PubMed] Conseil G et al: "Functional importance of three basic residues clustered at the cytosolic interface of transmembrane helix 15 in the multidrug and organic anion transporter MRP1 (ABCC1)."

No. Sentence Comment

23 Furthermore, the naturally occurring substitution of Arg433 by a Ser at the interface of CL4 and TM8 causes a selective decrease in LTC4 and estrone sulfate transport efficiency but increases drug resistance (23). Login to comment

PMID: 16330681 [PubMed] Wojnowski L et al: "NAD(P)H oxidase and multidrug resistance protein genetic polymorphisms are associated with doxorubicin-induced cardiotoxicity."

No. Sentence Comment

194 As opposed to polarized cells, in the cardiomyocytes MRP1 is found in the cytoplasm in addition to plasma membrane.37 It has been postulated that such an expression may permit sequestration of doxorubicin in lysosomes, ie, away from its target the nucleus.40 A low-frequency protein variant of MRP1 (Arg433Ser) has been shown to affect resistance to anthracyclines in vitro but has not been found in our study.41 The effect on doxorubicin resistance or transport of the variant 671Val, which shows an association with acute ACT in this study, has not been investigated.42 MRP2 has a substrate spectrum similar to that of MRP1 and increases the resistance to doxorubicin when overexpressed in HEK-293 cells.43 Physiologically, the MRP2 protein is expressed in the apical membrane of polarized cells in the liver and in the kidney, and there is no evidence for its expression in the heart. Login to comment
199 As opposed to polarized cells, in the cardiomyocytes MRP1 is found in the cytoplasm in addition to plasma membrane.37 It has been postulated that such an expression may permit sequestration of doxorubicin in lysosomes, ie, away from its target the nucleus.40 A low-frequency protein variant of MRP1 (Arg433Ser) has been shown to affect resistance to anthracyclines in vitro but has not been found in our study.41 The effect on doxorubicin resistance or transport of the variant 671Val, which shows an association with acute ACT in this study, has not been investigated.42 MRP2 has a substrate spectrum similar to that of MRP1 and increases the resistance to doxorubicin when overexpressed in HEK-293 cells.43 Physiologically, the MRP2 protein is expressed in the apical membrane of polarized cells in the liver and in the kidney, and there is no evidence for its expression in the heart. Login to comment

PMID: 16393888 [PubMed] Zhang J et al: "Metabolism and transport of oxazaphosphorines and the clinical implications."

No. Sentence Comment

627 Substitution of Arg433 with Ser predicted to be close to TM8 of MRP1 caused by the low frequency G1299T polymorphism in exon 10 leads to a substrate selective change in organic anion transport activity and drug resistance using MRP1-expressing HeLa cells (Conrad et al., 2002) or human leukemia CEM-7A cells (Assaraf et al., 2003). Login to comment

PMID: 16684361 [PubMed] Wang Z et al: "Nucleotide sequence analyses of the MRP1 gene in four populations suggest negative selection on its coding region."

No. Sentence Comment

8 SNPs E10/1299 G>T (R433S) and E16/2012 G>T (G671V) which occur at low frequency in only one or two of four populations examined were predicted to be functionally deleterious and hence are likely to be under negative selection. Login to comment
45 We found that SNPs E10/1299G>T, which resulted in arginine-serine substitution at amino acid position 433 (R433S) and E16/2012G>T, which resulted in glycine-valine substitution at amino acid position 671 (G671V), may potentially adversely affect the function of MRP1. Login to comment
123 2000 SNPe6* E10/1299 G>T R433S 0.12 benign -3.59003 0 0 0 1.39 Conrad et al . Login to comment

PMID: 17329911 [PubMed] Fukushima-Uesaka H et al: "Genetic variations and haplotype structures of the ABC transporter gene ABCC1 in a Japanese population."

No. Sentence Comment

26 In Caucasian populations, 1299GÀT (Arg433Ser), 1898GÀA (Arg633Gln), 2012GÀT (Gly671Val), and 4535CÀT (Ser1512Leu) have been reported.79) In addition, Arg433Ser decreases the transport activity for LTC4 and estrone sulfate, but not for estradiol 17b-glucuronide, in vitro.10) Ito et al. found 16 genetic polymorphisms, including 4 nonsynonymous and 8 synonymous ones, in 48 Japanese subjects.11) An in vitro functional study showed that one of the non-synonymous variations, 2168GÀA (Arg723Gln), leads to reduced transport activity for LTC4, estradiol 17b-glucuronide and methotrexate.12) However, no haplotype analysis has been reported for the Japanese population. Login to comment

PMID: 18220559 [PubMed] Yu XQ et al: "Multidrug resistance associated proteins as determining factors of pharmacokinetics and pharmacodynamics of drugs."

No. Sentence Comment

405 Important Single Nucleotide Polymorphisms (SNPs) of MRP Genes MRP Chromosomal location Amino acid variation Nucleotide variation Location References Cys43Ser Thr73Ile G128C C218T Exon2 Exon2 [239] Arg433Ser G1299T Exon10 [258] Gly671Val G2012T Exon16 [259] Arg723Gln G2168A Exon17 [239] MRP1 16p13.11-p13.12 Arg1058Gln G3173A Exon23 [239] C-24T Promoter [100, 239] Val417Ile G1249A Exon10 [100, 238, 239] Gly676Arg G2026C Exon16 [237] Try709Arg T2125C Exon17 [236] Arg768Trp Ser789Phe C2302T C2366T Exon18 Exon18 [100, 238, 239] I1173F R1150H A3517T G3449A Exon25 Exon25 [240] Ile1324Ile C3972T Exon28 [100, 239] MRP2 10q23-24 Ala1450Thr G4348A Exon31 [100, 238, 239] (Table 2) contd…. Login to comment

PMID: 19214144 [PubMed] Yin JY et al: "Characterization and analyses of multidrug resistance-associated protein 1 (MRP1/ABCC1) polymorphisms in Chinese population."

No. Sentence Comment

25 To date, only three naturally occurring mutations of MRP1/ABCC1 (Gly671Val, Arg433Ser, and Cys43Ser) have been fully investigated for their effects on MRP1/ ABCC1-mediated MDR [18-20]. Login to comment
28 For example, the substitution of highly conserved Arg433 by Ser significantly decreased the transport of leukotriene C4 and estrone sulfate, but increased doxorubicin resistance [18]. Login to comment
157 It has also been reported that Gly671Val and Arg433Ser mutations were found in Caucasian, but not in Asian populations [15,18,19]. Login to comment

PMID: 19687781 [PubMed] Siedlinski M et al: "ABCC1 polymorphisms contribute to level and decline of lung function in two population-based cohorts."

No. Sentence Comment

72 Table 2 Additive effects of 53 ABCC1 SNPs on the level and change of FEV1 in the Doetinchem cohort (n = 1152) Additive effect on the level of FEV1 (ml)a Additive effect on the change of FEV1 (ml/year)b SNP Position MAF (%) HWE P value B SE P B SE P rs4148330 50 region 34.2 0.96 - 63.2 20.4 0.002 - 1.8 1.4 0.20 rs504348 [28] 50 region 19.3 0.61 - 54.9 24.7 0.03 - 2.4 1.7 0.15 rs152022 Intron 1 20.3 0.12 9.0 24.0 0.71 0.4 1.6 0.83 rs215049 Intron 1 31.9 0.72 - 32.5 21.1 0.12 - 1.5 1.5 0.31 rs215068 Intron 1 30.9 0.21 22.7 21.5 0.29 - 0.7 1.5 0.64 rs215079 Intron 1 21.0 0.24 3.2 23.9 0.89 - 1.5 1.7 0.37 rs215100 Intron 1 32.1 0.77 - 48.8 20.9 0.02 - 1.0 1.4 0.50 rs246220 Intron 1 12.6 0.34 - 19.0 30.1 0.53 1.2 2.1 0.55 rs4781699 Intron 1 31.4 0.63 - 56.6 20.9 0.01 - 2.2 1.4 0.13 rs8060750 Intron 1 12.7 0.52 24.4 29.2 0.40 - 1.1 2.0 0.59 rs8045000 Intron 1 32.8 - - 51.2 20.8 0.01 - 1.8 1.4 0.20 rs7190484 Intron 1 38.1 - - 48.8 20.1 0.02 - 1.8 1.4 0.19 rs7196970 Intron 1 40.6 - - 28.6 20.1 0.15 - 1.0 1.4 0.49 rs215101 Intron 1 10.5 - 36.2 33.1 0.27 - 0.9 2.2 0.70 rs4781711 Intron 1 6.9 - - 22.0 41.5 0.60 0.3 2.9 0.91 rs215059 Intron 1 14.2 - 15.6 28.8 0.59 0.8 2.0 0.69 rs4148337 Intron 3 31.9 0.72 - 25.4 21.0 0.23 - 1.0 1.4 0.51 rs4781718 Intron 3 14.3 0.39 - 0.1 27.8 0.99 - 2.1 1.9 0.28 rs246213 Intron 5 8.8 0.97 - 46.9 34.9 0.18 - 2.9 2.4 0.24 rs246240 Intron 5 18.0 0.28 3.1 25.3 0.90 - 0.7 1.7 0.71 rs7188937 Intron 5 46.4 - - 22.0 19.2 0.25 - 1.9 1.3 0.14 rs246233 Intron 6 9.0 - - 8.8 37.7 0.82 1.8 2.6 0.49 rs246230 Intron 7 14.6 0.42 - 5.2 27.5 0.85 - 0.1 1.9 0.97 rs193537 Intron 7 29.5 - - 14.0 22.3 0.53 - 0.2 1.5 0.89 rs60782127 (Arg433Ser) [29,30] Exon 10 2.2 1.00 - 25.2 68.0 0.71 3.3 4.7 0.48 rs12445600 Intron 10 27.8 0.57 - 22.8 22.0 0.30 - 0.5 1.5 0.74 rs4148344 Intron 10 15.7 - 12.5 28.4 0.66 0.5 1.9 0.80 rs35596 Intron 12 23.1 0.44 - 11.0 23.7 0.64 0.9 1.6 0.57 rs35597 Intron 12 46.7 0.33 32.9 20.1 0.10 3.1 1.4 0.02 rs35610 Intron 13 15.8 0.30 - 38.0 27.5 0.17 - 1.6 1.9 0.41 rs35621 Intron 14 10.7 0.61 -7.8 31.4 0.80 - 3.0 2.1 0.17 rs4148353 Intron 15 14.8 0.46 - 4.5 28.3 0.87 0.4 2.0 0.84 rs45511401 (Gly671Val) [30,31] Exon 16 4.9 1.00 22.5 45.4 0.62 - 0.7 3.1 0.81 rs2074086 Intron 18 31.9 0.43 11.7 21.0 0.58 - 1.0 1.4 0.49 rs2074087 Intron 18 14.2 - 42.1 30.6 0.17 - 3.6 2.1 0.08 rs4148358 Intron 19 23.5 0.94 11.1 23.2 0.63 - 0.1 1.6 0.93 rs2283515 Intron 19 46.6 - - 17.5 19.8 0.38 - 2.8 1.3 0.04 rs12449239 Intron 19 14.9 - - 20.4 29.8 0.49 - 0.7 2.0 0.74 rs11864374 Intron 21 22.6 0.62 - 27.4 24.2 0.26 - 2.9 1.6 0.08 rs9933511 Intron 21 27.1 0.09 9.5 22.7 0.68 - 0.5 1.5 0.75 rs3819552 Intron 21 49.0 0.57 6.6 19.9 0.74 2.7 1.4 0.04 rs2238475 Intron 23 4.6 0.16 - 10.8 46.0 0.82 1.2 3.2 0.71 rs2238476 Intron 23 5.9 0.74 28.0 41.5 0.50 - 1.5 2.8 0.59 rs12599429 Intron 26 31.8 0.85 17.7 21.5 0.41 2.0 1.5 0.17 rs212079 Intron 26 4.1 0.85 - 20.3 50.0 0.69 - 5.9 3.4 0.09 rs212083 Intron 27 19.5 0.46 - 10.6 24.6 0.67 - 3.9 1.7 0.02 rs2230671 (Ser1334Ser) Exon 28 29.8 0.98 5.7 21.6 0.79 1.9 1.5 0.19 rs3743527 30 UTR 22.5 0.09 - 58.6 23.5 0.01 - 1.3 1.6 0.44 rs212090 30 UTR 45.5 - - 11.2 19.4 0.56 4.3 1.3 0.001 rs212093 30 region 45.3 0.10 - 42.5 19.3 0.03 - 2.9 1.3 0.03 rs4148382 30 region 9.7 1.00 94.2 33.4 0.005 - 3.6 2.3 0.11 rs212094 30 region 18.0 - 15.9 25.8 0.54 - 5.3 1.8 0.003 rs12448760 30 region 29.1 - -7.5 21.6 0.73 2.7 1.5 0.07 The lack of HWE P values indicates haplotype-tagged SNPs. Login to comment

PMID: 21143116 [PubMed] He SM et al: "Structural and functional properties of human multidrug resistance protein 1 (MRP1/ABCC1)."

No. Sentence Comment

735 The naturally occurring replacement of Arg433 by Ser (neutral) at the interface of CL4 and TM8 caused a 2-fold decrease in LTC4 and estrone sulfate transport but were 2.1-fold more resistant to doxorubicin while while resistance to etoposide (VP-16) and vincristine remained unchanged [363]. Login to comment
816 There are at least 15 naturally occurring mutations identified in MRP1/ABCC1, including Cys43Ser in TM1, Thr73Ile in CL1, Ser92Phe in TM2, Arg230Asn in L0, Val353Met at TM6/TM7 interface, Arg433Ser in TM8, Gly671Val in TM11, Arg723Gln located between the Walker A and Walker B motifs of NBD1, Ala861Thr at NBD1/TM12 interface, Ala989Thr in TM12, Cys1047Ser in TM13, Arg1058Gln in CL7, Val1146Ile in CL7, Thr1337Ala between the Walker A and Walker B motifs of NBD2, and Thr1401Met, and many of them have been found to affect its transport activity [171, 362, 363, 366, 367, 377-384]. Login to comment
819 A naturally-occurring mutation of 1299G>T in exon 10 causing Arg433Ser in TM8 of TMD1 resulted in decreased transport of LTC4 and increased resistance to doxorubicin [363]. Login to comment

PMID: 21182225 [PubMed] Zhao J et al: "Promoter polymorphism of MRP1 associated with reduced survival in hepatocellular carcinoma."

No. Sentence Comment

46 G1299T (Arg433Ser) confers resistance to doxorubicin by reducing intracellular drug accumulation in HeLa cells that stably express mutant MRP1, whereas the G3173A (Arg1058Gln) variation increases the response to etoposide in HEK293 and CHO-K1 cells[12,13] . Login to comment

PMID: 21736601 [PubMed] Yin JY et al: "ABCC1 polymorphism Arg723Gln (2168G > A) is associated with lung cancer susceptibility in a Chinese population."

No. Sentence Comment

24 For example, Cys43Ser (128G>C), which is located in the NH2 proximal region, was found to be important for the maintenance of MRP1/ABCC1-induced drug resistance.6 Another polymorphism, Arg433Ser (1299G>A), has the potential to increase resistance to doxorubicin in transfected Hela cells.7 Our previous investigation also showed that Arg723Gln (2168G>A) could significantly reduce MRP1/ABCC1 induced MDR.8 Furthermore, a number of in vivo studies have provided the clinical data to support the important role of MRP1/ABCC1 polymorphisms in disease susceptibility, drug metabolism and toxicity. Login to comment

PMID: 14598019 [PubMed] Gerloff T et al: "Impact of genetic polymorphisms in transmembrane carrier-systems on drug and xenobiotic distribution."

No. Sentence Comment

166 Digoxin plasma levels are inversely correlated with Pgp expression levels The functional relevance of the MRP1 SNPs G671 V, positioned within NBD1, and of Arg433Ser, located in a putative cytoplasmic loop, was tested using in vitro transport assays (Conrad et al. 2002). Login to comment
167 While the first did not change MRP1 transport activity for conjugated organic anions, the Arg433Ser polymorphism reduced transport activity of the substrates leucotriene C4 and estrone sulfate by half due to a decrease in Vmax for both compounds, whereas Km remained unchanged. Login to comment
168 In addition, doxorubicin-resistance of cells overexpressing the mutant Arg433Ser MRP1 was 2-fold higher compared to the wild-type. Login to comment

PMID: 20103563 [PubMed] Klaassen CD et al: "Xenobiotic, bile acid, and cholesterol transporters: function and regulation."

No. Sentence Comment

7118 Nucleotide Change Amino Acid Change In Vitro Function Protein Expression/Localization ABCC1 MRP1 G128C C43S 1↔ Intracellular C218T T73I 1↔ Normal C257T S92F 2↔ Normal C350T T117M 2↔ Normal G689A R230Q ↔ Normal G1057A V353M N.D. N.D. G1299T R433S 2↔ Normal G1898A R633Q 2↔ Normal G2012T G671V ↔ Normal G2168A R723Q 2 Normal G2965A A989T 2↔ Normal G3140C C1047S 1↔ Normal G3173A R1058Q ↔ Normal C4535T S1512L ↔ Normal ABCC2 MRP2 C-24T N.D. N.D. G1058A R353H N.D. N.D. G1249A V417I ↔ Normal C2366T S789F 12 Intracellular T2780G L927R N.D. N.D. C3298T R1100C N.D. N.D. G3299A R1100H N.D. N.D. T3563A V1188E N.D. N.D. G4348A A1450T ↔ Normal/Intracellular G4544A C1515Y N.D. N.D. ABCC3 MRP3 G32A G11D ↔ Normal C202T H68Y N.D. N.D. G296A R99Q N.D. Normal C1037T S346F 2 Normal C1537A Q513K N.D. N.D. T1643A L548Q N.D. N.D. G1820A S607N 2 Normal C2221T Gln741STOP N.D. N.D. G2293C V765L ↔ Normal G2395A V799M N.D. N.D. C2758T P920S 1 Normal G2768A R923Q 1 Normal C3657A S1219R N.D. N.D. C3856G R1286G ↔ Normal G3890A R1297H N.D. N.D. C4042T R1348C 1 Normal A4094G Q1365R ↔ Normal C4141A R1381S ↔ Intracellular C4217T T1406M N.D. N.D. G4267A G1423R N.D. N.D. ABCC4 MRP4 C52A L18I N.D. N.D. C232G P78A 2↔ Normal T551C M184T N.D. N.D. G559T G187W 2 Reduced A877G K293E ↔ Normal G912T K304N ↔ Normal C1067T T356M N.D. N.D. C1208T P403L 2↔ Normal G1460A G487E 2 Normal A1492G K498E ↔ Normal A1875G I625M N.D. N.D. C2000T P667L N.D. N.D. A2230G M744V ↔ Normal G2269A E757K N.D. Intracellular G2459T R820I N.D. N.D. G2560T V854F N.D. N.D. G2698T V900L N.D. N.D. G2867C C956S 1↔ Normal G3211A V1071I ↔ Normal C3425T T1142M N.D. N.D. G3659A R1220Q N.D. N.D. A3941G Q1314R N.D. N.D. 2, reduced function; 1, increased function; ↔, no change in function; N.D. not determined. Login to comment
7115 Nucleotide Change Amino Acid Change In Vitro Function Protein Expression/Localization ABCC1 MRP1 G128C C43S 1࢒ Intracellular C218T T73I 1࢒ Normal C257T S92F 2࢒ Normal C350T T117M 2࢒ Normal G689A R230Q ࢒ Normal G1057A V353M N.D. N.D. G1299T R433S 2࢒ Normal G1898A R633Q 2࢒ Normal G2012T G671V ࢒ Normal G2168A R723Q 2 Normal G2965A A989T 2࢒ Normal G3140C C1047S 1࢒ Normal G3173A R1058Q ࢒ Normal C4535T S1512L ࢒ Normal ABCC2 MRP2 C-24T N.D. N.D. G1058A R353H N.D. N.D. G1249A V417I ࢒ Normal C2366T S789F 12 Intracellular T2780G L927R N.D. N.D. C3298T R1100C N.D. N.D. G3299A R1100H N.D. N.D. T3563A V1188E N.D. N.D. G4348A A1450T ࢒ Normal/Intracellular G4544A C1515Y N.D. N.D. ABCC3 MRP3 G32A G11D ࢒ Normal C202T H68Y N.D. N.D. G296A R99Q N.D. Normal C1037T S346F 2 Normal C1537A Q513K N.D. N.D. T1643A L548Q N.D. N.D. G1820A S607N 2 Normal C2221T Gln741STOP N.D. N.D. G2293C V765L ࢒ Normal G2395A V799M N.D. N.D. C2758T P920S 1 Normal G2768A R923Q 1 Normal C3657A S1219R N.D. N.D. C3856G R1286G ࢒ Normal G3890A R1297H N.D. N.D. C4042T R1348C 1 Normal A4094G Q1365R ࢒ Normal C4141A R1381S ࢒ Intracellular C4217T T1406M N.D. N.D. G4267A G1423R N.D. N.D. ABCC4 MRP4 C52A L18I N.D. N.D. C232G P78A 2࢒ Normal T551C M184T N.D. N.D. G559T G187W 2 Reduced A877G K293E ࢒ Normal G912T K304N ࢒ Normal C1067T T356M N.D. N.D. C1208T P403L 2࢒ Normal G1460A G487E 2 Normal A1492G K498E ࢒ Normal A1875G I625M N.D. N.D. C2000T P667L N.D. N.D. A2230G M744V ࢒ Normal G2269A E757K N.D. Intracellular G2459T R820I N.D. N.D. G2560T V854F N.D. N.D. G2698T V900L N.D. N.D. G2867C C956S 1࢒ Normal G3211A V1071I ࢒ Normal C3425T T1142M N.D. N.D. G3659A R1220Q N.D. N.D. A3941G Q1314R N.D. N.D. 2, reduced function; 1, increased function; ࢒, no change in function; N.D. not determined. Login to comment

PMID: 22565165 [PubMed] Grover S et al: "Genetic association analysis of transporters identifies ABCC2 loci for seizure control in women with epilepsy on first-line antiepileptic drugs."

No. Sentence Comment

87 - 330-21247T > C Intron 1 0.005 6 rs4148731 chr7:87239329 c.-330 - 8935C > T Intron 1 0.000 7 rs9282564 chr7:87229440 c.61A > G Exon 3 (Asn21Asp) 0.000 8 rs9282565 chr7:87214875 c.239C > A Exon 5 (Ala80Glu) 0.000 9 rs28381826 chr7:87214531 c.286 + 297G > A Intron 5 0.000 10 rs1989830 chr7:87205663 c.287 - 6124C > T Intron 5 0.135 11 rs2520464 chr7:87201086 c.287 - 1547A > G Intron 5 0.409 12 rs2235023 chr7:87190452 c.827+ 127G > A Intron 9 0.000 13 rs10276036 chr7:87180198 c.1000 - 44C > T Intron 10 0.401 14 rs2229109 chr7:87179809 c.1199G > A Exon 12 (Ser400Asn) 0.000 15 rs1128503 chr7:87179601 c.1236T > C Exon 13 (Gly412Gly) 0.390 16 rs2235036 chr7:87175271 c.1795G > A Exon 16 (Ala599Thr) 0.000 17 rs2235039 chr7:87165854 c.2401G > A Exon 21 (Val801Met) 0.000 18 rs2235040 chr7:87165750 c.2481 + 24G > A Intron 21 0.155 19 rs2032581 chr7:87160810 c.2485A > G Exon 22 (Ile829Val) 0.000 20 rs2032582 chr7:87160618 c.2677T/A > G Exon 22 (Ser/Thr893Ala) 0.318 21 rs7779562 chr7:87144816 c.3085 -72G > C Intron 25 0.043 22 rs2707944 chr7:87144641 c.3188C > G Exon 26 (Ala1063Gly) 0.000 23 rs2229107 chr7:87138659 c.3421A > T Exon 27 (Thr1141Ser) 0.000 24 rs1045642 chr7:87138645 c.3435T > C Exon 27 (Ile1145Ile) m Expression and activity [28] m mRNA expression [29] Altered substrate specificity [30] 0.375 25 rs2235048 chr7:87138511 c.3489 + 80C > T Intron 27 0.381 26 rs17064 chr7:87133470 c.3932A > T 30 UTR 0.000 ABCC1 1 rs504348 chr16:16043174 rs50438C > G Near gene region k Promoter activity [31] 0.135 2 rs215106 chr16:16047542 c.48 + 3886A > G Intron 1 0.210 3 rs215049 chr16:16070768 c.48 + 27112G > C Intron 1 0.245 4 rs246220 chr16:16082128 c.49 - 19545C > G Intron 1 0.118 5 rs119774 chr16:16086833 c.49 - 14840G > A Intron 1 0.089 6 rs246217 chr16:16090354 c.49 - 11319C > A Intron 1 0.118 7 rs2014800 chr16:16099966 c.49 - 1707C > T Intron 1 0.398 8 rs41494447 chr16:16101842 c.218C > T Exon 2 (Thr73Ile) 0.000 9 rs4781712 chr16:16103232 c.226 - 401A > G Intron 2 0.355 10 rs246240 chr16:16119024 c.616 -7942A > G Intron 5 0.114 11 rs924135 chr16:16123459 c.616 - 3507A > T Intron 5 0.412 12 rs903880 chr16:16130514 c.809 + 54C > A Intron 7 0.147 13 rs8187852 chr16:16139709 c.1057G > A Exon 9 (Met353Val) 0.000 14 rs35587 chr16:16139714 c.1062T > C Exon 9 (Asn354Asn) 0.182 15 rs35592 chr16:16141823 c.1219 - 176T > C Intron 9 0.172 16 rs60782127 chr16:16142079 c.1299G > T Exon 10 (Arg433Ser) k Transport of leukotriene C4 and estrone sulfate [32] 0.008 17 rs3765129 chr16:16149901 c.1474 - 48C > T Intron 11 0.032 18 rs35597 chr16:16158034 c.1678 - 3979G > A Intron 12 0.320 19 rs35621 chr16:16168608 c.1913 - 1575C > T Intron 14 0.103 20 rs45511401 chr16:16173232 c.2012G > T Exon 16 (Gly671Val) 0.024 21 rs4148356 chr16:16177275 c.2168G > A Exon 17 (Arg723Gln) 0.000 22 rs3851713 chr16:16184873 c.2644 + 428A > T Intron 19 0.340 23 rs2239995 chr16:16192565 c.2645 - 3919G > A Intron 19 0.324 24 rs11864374 chr16:16201885 c.2871 + 1155G > A Intron 21 0.338 25 rs35529209 chr16:16205325 c.2965G > A Exon 22 (Thr989Ala) k Transport of estradiol 17b-glucuronide [32] 0.000 26 rs3887893 chr16:16205501 c.3079 + 62G > A Intron 22 0.448 27 rs13337489 chr16:16208683 c.3140G > C Exon 23 (Ser1047Cys) 0.000 28 rs2299670 chr16:16220858 c.3819 + 1090A > G Intron 26 0.399 29 rs8057331 chr16:16230411 c.4202C > T Exon 29 (Thr1401Met) 0.000 30 rs212090 chr16:16236004 c.5462T > A 30 UTR 0.357 31 rs212093 chr16:16237754 rs212093G > A Near gene region 0.429 32 rs4148382 chr16:16238494 rs4148382G > A Near gene region 0.034 ABCC2 1 g.-1774G > delG chr10:101535688 g.-1774G > delG Near gene region k Promoter activity [33] 0.000 2 rs1885301 chr10:101541053 c.-1549G > A Near gene region k Promoter activity [haplotype containing (- 1549A)-(- 24T)] [33] k Clearance of irinotecan (ABCC2*2 containing the G allele) [34] 0.379 450 Pharmacogenetics and Genomics 2012, Vol 22 No 6 Table 2 (continued) N dbSNP ida Positionb Allelesc Gene location (effect) Function MAF 3 rs2804402 chr10:101541583 c. Login to comment

PMID: 15083066 [PubMed] Lee YM et al: "Identification and functional characterization of the natural variant MRP3-Arg1297His of human multidrug resistance protein 3 (MRP3/ABCC3)."

No. Sentence Comment

209 However, a naturally occurring variant (MRP1-Arg433 Ser) has been described, which was correctly routed to the plasma membrane but showed a selective decrease in organic anion transport [45]. Login to comment

PMID: 17412754 [PubMed] Wang Z et al: "Signatures of recent positive selection at the ATP-binding cassette drug transporter superfamily gene loci."

No. Sentence Comment

184 Nonetheless, two very low frequency (,2%) non-synonymous ABCC1 SNPs, e22/G2965A (Ala989Thr) and e10/G1299GT (Arg433Ser), were reported experimentally to result in decreased estradiol 17b-glucoronide (53) and organic anion transport but increased doxorubicin resistance (54), respectively. Login to comment
183 Nonetheless, two very low frequency (,2%) non-synonymous ABCC1 SNPs, e22/G2965A (Ala989Thr) and e10/G1299GT (Arg433Ser), were reported experimentally to result in decreased estradiol 17b-glucoronide (53) and organic anion transport but increased doxorubicin resistance (54), respectively. Login to comment

PMID: 18377430 [PubMed] Lal S et al: "Influence of ABCB1 and ABCG2 polymorphisms on doxorubicin disposition in Asian breast cancer patients."

No. Sentence Comment

161 (44) Naturally occurring non-synonymous mutations (Arg433Ser) in the MRP1 cytoplasmic domain, leading to doxorubicin resistance,(45) also needs further investigation in terms of its role in altering doxorubicin disposition in cancer patients. Login to comment

PMID: 16815813 [PubMed] Choudhuri S et al: "Structure, function, expression, genomic organization, and single nucleotide polymorphisms of human ABCB1 (MDR1), ABCC (MRP), and ABCG2 (BCRP) efflux transporters."

No. Sentence Comment

311 Another SNP G2012T in exon 16 (Gly671Val) was found to have no functional consequences, whereas exon 10 SNP G1299T (Arg433Ser; in the cytosolic interface of transmembrane segment 8) resulted in significant decrease in the transport ability of many organic anions but increased doxorubicin resistance (Conseil, Deeley, and Cole 2005). Login to comment

PMID: 22293538 [PubMed] Jungsuwadee P et al: "The G671V variant of MRP1/ABCC1 links doxorubicin-induced acute cardiac toxicity to disposition of the glutathione conjugate of 4-hydroxy-2-trans-nonenal."

No. Sentence Comment

3 Methods We established stable HEK293 cell lines overexpressing wild-type MRP1 (HEKMRP1), G671V (HEKG671V), and R433S (HEKR433S), a variant not associated with doxorubicin-induced cardiotoxicity and investigated the sensitivity of HEKG671V cells to doxorubicin and transport capacity of G671V toward GS-HNE. Login to comment
4 Results In ATP-dependent transport studies using plasma membrane-derived vesicles, the Vmax (pmol/min/mg) for GS-HNE transport was the lowest for G671V (69 ± 4) and the highest for R433S (972 ± 213) compared with wild-type MRP1 (416 ± 22), whereas the Km values were 2.8 ± 0.4, 6.0 or more, and 1.7 ± 0.2 lmol/l, respectively. Login to comment
20 Among several variants, they identified Arg433Ser (R433S), Original article 273 1744-6872 c 2012 Wolters Kluwer Health | Lippincott Williams & Wilkins DOI: 10.1097/FPC.b013e328350e270 located in the second transmembrane spanning domain, and Gly671Val (G671V; rs45511401), near the Walker A motif in the first nucleotide-binding domain. Login to comment
21 The R433S MRP1 variant showed a 50% decreased transport maximum for leukotriene C4 (LTC4) [8]; however, cells expressing this variant were more resistant to doxorubicin than those expressing wild-type (WT) MRP1 [8]. Login to comment
37 In this study, we examined whether cells expressing the G671V and R433S variants versus WT MRP1 were more sensitive to doxorubicin, and characterized the transport properties of WT MRP1 relative to the G671V and R433S MRP1 variants, specifically with respect to GS-HNE transport activity. Login to comment
46 MRP1 variants of G671Vand R433S were generated using the QuickChange II XL site-directed mutagenesis kit (Stratagene, La Jolla, California, USA) according to the manufacturer`s instructions with the following mutagenic primers (for G671V, forward primer: 50 -TCCATCCCCGAAGTTGCTTTGGTG 274 GCCGTG-30 and reverse primer: 50 -CACGGCCACCAAAG CAACTTCGGGGATGGA-30 ; for R433S, forward primer: 50 -GTGGACGCTCAGAGCTTCATGGACTTGGC-30 and reverse primer: 50 -GCCAAGTCCATGAAGCTCTGAGCGT CCAC-30 ). Login to comment
114 In an attempt to understand this phenomenon, we developed HEK293 cell lines that stably expressed human MRP1 SNPs G671V, and three different control cells: HEK293 cells transfected with pUSEamp(+) vector (HEKpUSE), WT MRP1 (HEKMRP1), or the R433S variant (HEKR433S), which is not associated with doxorubicin-induced acute cardiac toxicity. Login to comment
141 MRP1 G671V decreases GS-HNE transport Jungsuwadee et al. 277 Fig. 1 (b) 3000 MRP1pUSE G671V R433S *** *** *** 2000 Meanfluorescenceintensity 1000 0 pUSE MRP1 G671V R433S (a) (c) 10.0 * * * 7.5 5.0 2.5 MRP1/18S 0.0 pUSE MRP1 G671V R433S Expression in stable cell lines of multidrug resistance-associated protein 1 (MRP1) and its variants. Login to comment
143 Plasmids containing wild-type MRP1 or the MRP1 variants G761V and R433S were transfected into HEK293 cells to generate stable cell lines. Login to comment
149 Fig. 2 (a) (b) Genotype Doxorubicin IC50 nmol/l (95% CI) pUSE 58.0 (51.4-65.5) MRP1 463.0 (332.7-644.2) G671V 181.4 (137.4-239.5) R433S 645.2 (158.3-2630.0) pUSE125 100 75 50 Percentsurvival 25 0 MRP1 G671V R433S 10 Doxorubicin (μmol/l) -1-2-3 Percent survival of HEKpUSE, HEKMRP1, HEKR433S, and HEKG671V cells in the presence of doxorubicin. Login to comment
156 The G671V variant was comparable with WT MRP1 with respect to LTC4 transport, whereas LTC4 transport was decreased by 75% in the R433S variant compared with WT MRP1 (Fig. 6b), consistent with previous reports [7,8]. Login to comment
160 In contrast, the Vmax of the R433S variant was increased over two-fold relative to WT MRP1. Login to comment
162 The estimate of the Km for the R433S variant was higher (Z 6 mmol/l), consistent with its increased Vmax (Fig. 6c). Login to comment
165 To investigate whether the cardiac sarcolemma can transport GS-HNE, and the Fig. 3 (a) (b) (c) (d) 1.5 ** ** **** ** ** ** *** ** 1.0 Fractionofcontrol doxorubicinaccumulation Fractionofcontrol doxorubicinaccumulation 0.5 0.0 8 250 200 150 100 GS-HNE(%control) 50 0 pUSE MRP1 G671V R433S 6 4 GS-HNE(nmol/mg) 2 0 Doxorubicin (1.5 μmol/l) - + - + - + - + pUSE MRP1 G671V R433S 1.5 1.0 0.5 0.0 pUSE 1 h M RP1 h G 671V 1 h R433S 1 h pUSE 24 h M RP1 24 h G 671V 24 h R433S 24 h Doxorubicin and glutathione-conjugated 4-hydroxy-2-trans-nonenal (GS-HNE) retention in HEKpUSE, HEKMRP1, HEKR433S, and HEKG671V cells. Login to comment
182 These data are in contrast with an earlier report [8] showing that HeLa cells expressing R433S are two-fold more resistant than cells expressing WT MRP1. Login to comment
185 We next examined cellular GS-HNE levels after treatment of cells with 1.5 mmol/l of doxorubicin for 24 h and Fig. 4 20(a) (b) (c) 15 12.5 10.0 7.5 5.0Glutathione (nmol/mgprotein) GSSG(nmol/mgprotein) 2.5 0.0 15 10 5 Glutathione/GSSGratio 0 pUSE pUSE 0.8 *** *** *** ** *** ***0.6 0.4 0.2 0.0 M RP1 G 671V R433S M RP1 G 671V R433S pUSE M RP1 G 671V R433S 10 Glutathione(nmol/mg) Glutathione/GSSGratio 5 0 0 16 14 12 10 8 0 10 20 30 40 50 Time (h) 25 50 Time (h) 1.5 1.0 0.5 0.0 0 10 20 30 40 50 Time (h) GSSG(nmol/mg) Time course of intracellular glutathione and GSSG in HEKpUSE, HEKMRP1, HEKR433S, and HEKG671V cells. Login to comment
205 It is also interesting to note that the R433S variant showed significantly increased transport of GS-HNE, despite decreased transport of LTC4 (Fig. 6b) and estrone sulfate, and unaltered transport of E217G [8]. Login to comment
219 Although increased retention of doxorubicin could also contribute to the increased oxidative stress in cells expressing the G671V variant, because amino acids in the third membrane spanning domain, especially between amino acids 959 and 1187, are considered most critical for doxorubicin transport [35], it seems less Fig. 6 Genotype Km ± SE (μmol/l) Vmax± SE (pmol/min/mg) Vmax/Km (mg/l/min) pUSE (95% CI) 6.3 ±12.9 (0.0-33.5) 28.4± 36.4 (0.0-104.8) 4.5 MRP1 (95% CI) 1.7 ±0.2 (1.2-2.1) 416.1±21.7 (371.1-461.2) 244.7 G671V (95% CI) 2.8± 0.4 (2.1-3.6) 69.1 ±4.1 (60.5-77.7) 24.7 R433S (95% CI) ≥6 972.1± 212.7 (531.0-1413) Approximately 160 pUSE MRP1 600 500 Protein(pmol/min/mg) 400 300 200 100 0 0 1 2 3 GS-HNE (μmol/l) 4 5 6 7 G671V R433S 125 *** *** 100 75 50 25 0 PercentLTC4efflux relativetoWT M RP1G 671VR433S M RP1+glutathione 5 G 671V+glutathione 5 R433S+glutathione 5 M RP1+glutathione 0.5 G 671V+glutathione 0.5 R433S+glutathione 0.5 2.5 (a) (b) (c) MRP1 pUSE/293 MRP1 G671V R433S Na/K-ATPase MRP1/NaK-ATPase banddensity (arbitraryunit) 2.0 1.5 1.0 0.5 0.0 MRP1 G671V R433S Multidrug resistance-associated protein 1 (MRP1) expression and transport activities of HEKMRP1, HEKR433S, and HEKG671V. Login to comment

PMID: 20367109 [PubMed] Giraud C et al: "ABC transporters in human lymphocytes: expression, activity and role, modulating factors and consequences for antiretroviral therapies."

No. Sentence Comment

179 3.1.2.1 ABCC1/MRP1 In 2001, Conrad et al. described two non-synonymous polymorphisms, R433S and G671V. Login to comment

PMID: 24080162 [PubMed] Conseil G et al: "Two polymorphic variants of ABCC1 selectively alter drug resistance and inhibitor sensitivity of the multidrug and organic anion transporter multidrug resistance protein 1."

No. Sentence Comment

27 Previously, we generated and partially characterized recombinant forms of ABCC1 nsSNPs: rs45511401 (2012G.T; G671V), rs60782127 (1299G.T; R433S), and rs41395947 (128G.C; C43S) (Conrad et al., 2001, 2002; Leslie et al., 2003). Login to comment
29 In contrast, the R433S mutant exhibited reduced LTC4 and estrone sulfate transport levels (Conrad et al., 2002). Login to comment
30 Cells expressing MPR1-R433S also showed increased doxorubicin resistance, suggesting R433S can better efflux this drug despite its lower organic anion transport activity. Login to comment
34 In contrast to our experimental data, the predictive algorithms SIFT (Sorting Tolerant From Intolerant) and PolyPhen indicated that G671V would adversely affect MRP1 function but C43S and A989T were less likely to do so, whereas predictions for R433S were mixed (L&#e9;tourneau et al., 2005). Login to comment

PMID: 24670052 [PubMed] Kunicka T et al: "Importance of ABCC1 for cancer therapy and prognosis."

No. Sentence Comment

148 However, substitution of highly conserved Arg433 with serine (G1299T, rs60782127) significantly reduces transport of estrone-3-sulfate and LTC4 and increases resistance to doxorubicin (Conrad et al., 2002). Login to comment
150 Interestingly, this Arg433Ser substitution was associated with increased doxorubicin resistance in transfected cells. Login to comment
159 NCBI ID Reference Amino acid exchange Nucleotide exchange Location Function MAFa rs41395947 Cys43Ser G128C Exon 2 Non-synonymous Unknown rs41494447 Thr73Ile C218T Exon 2 Non-synonymous T &#bc; 0.003 rs8187844 Ser92Phe C257T Exon 3 Non-synonymous T &#bc; 0.004 rs8187848 Arg230Gln G689A Exon 7 Non-synonymous A &#bc; 0.009 rs2230669 Pro272Pro G816A Exon 8 Synonymous A &#bc; 0.037 rs246221 Val275Val T825C Exon 8 Synonymous C &#bc; 0.301 rs35592 non-coding T-176C Intron 9 Non-coding C &#bc; 0.257 rs60782127 Arg433Ser G1299T Exon 10 Non-synonymous T &#bc; 0.004 rs35605 Leu562Leu T1684C Exon 13 Synonymous T &#bc; 0.173 rs112282109 Arg633Gln G1898A Exon 14 Non-synonymous A &#bc; 0.004 rs45511401 Gly671Val G2012T Exon 16 Non-synonymous T &#bc; 0.050 rs4148356 Arg723Gln G2168A Exon17 Non-synonymous A &#bc; 0.027 rs35529209 Ala989Thr G2965A Exon 22 Non-synonymous Unknown rs13337489 Cys1047Ser G3140C Exon 23 Non-synonymous C &#bc; 0.000 rs41410450 Arg1058Gln G3173A Exon 23 Non-synonymous Unknown rs2238476 non-coding G-1960A Intron 23 Non-coding T &#bc; 0.062 rs2230671 Ser1334Ser G4002A Exon 28 Synonymous T &#bc; 0.208 rs28364006 Thr1337Ala A4009G Exon 28 Non-synonymous Unknown rs369410659 Ser1512Leu C4535T Exon 31 Non-synonymous Unknown a Minor allele frequencies for Caucasinans in dbSNP based on HapMap-CEU population or 1000 genomes. Login to comment

PMID: 26395522 [PubMed] Slomka M et al: "Genetic variation of the ABC transporter gene ABCC1 (Multidrug resistance protein 1-MRP1) in the Polish population."

No. Sentence Comment

136 Variants c.596C > T (p.Ser199Leu) and c.814C > T (p.Pro272Ser) were located in the third intracellular loop (between TM5 and TM6), variant c.1299G > T (p.Arg433Ser) in the fourth intracellular loop, variant c.3196C > T (p.Arg1066Trp) in the seventh intracellular loop (between TM7 and TM8). Login to comment
142 The novel variant c.596C > T (p.Ser199Leu) was estimated as a probably damaging substitution, likewise as four others: c.1299G > T (Arg433 Ser), c.2012G > T (p.Gly671Val), c.3886C > T (p.Arg 1296Trp) and c.3901C > T (p.Arg1301Cys). Login to comment
143 On the other hand, analysis for HumVar-trained model indicated that three polymorphisms: c.1299G > T (Arg433Ser), c.2012G > T (p.Gly671Val), c.3901C > T (p.Arg1301Cys), lead to probably damaging substitutions and two others, c.596C > T (p.Ser199Leu) and c.3886C > T (p.Arg1296Cys), are possibly damaging variants. Login to comment
144 Table 2 Summary of ABCC1 variants detected during scanning by HRM Exon scanned by HRM dbSNP ID Variant position NM_004996.3: Intron/amino acid residue NP_004987.2: Observed genotypesa, b (n) HWE exact test P-valuec MAFd R/R R/V V/V 2 rs8187843 c.225 + 26G > A Intron 164 25 0 1 (A) 0.066 4 rs587783373* c.352-79G > A Intron 185 1 0 1 (A) 0.003 4 rs4148337 c.352-66 T > C Intron 15 80 91 0.727 (T) 0.296 5 rs483352860* c.596C > T p.Ser199Leu 186 1 0 1 (T) 0.003 6 rs8187846 c.677 + 17C > T Intron 188 1 0 1 (T) 0.003 7 rs483352864* c.809 + 16C > T Intron 188 1 0 1 (T) 0.003 7 rs45609533 c.809 + 31G > T Intron 183 5 0 1 (T) 0.013 7 rs903880 c.809 + 54C > A Intron 112 65 11 0.684 (A) 0.231 7 rs246232 c.809 + 64C > G Intron 84 90 14 0.174 (G) 0.314 8 rs546943313 c.810-73C > T Intron 187 1 0 1 (T) 0.003 8 rs200194736 c.814C > T p.Pro272Ser 187 1 0 1 (T) 0.003 8 rs2230669 c.816G > A p.Pro272= 172 16 0 1 (A) 0.043 8 rs246221 c.825 T > C p.Val275= 84 92 12 0.059 (C) 0.309 8 rs587783372* c.855G > A p.Pro285= 187 1 0 1 (A) 0.003 9 rs35587 c.1062 T > C p.Asn354= 78 91 16 0.185 (C) 0.332 9 rs35588 c.1218 + 8A > G Intron 82 91 16 0.245 (G) 0.327 9 rs483352877* c.1218 + 9C > T Intron 188 1 0 1 (T) 0.003 10 rs60782127 c.1299G > T p.Arg433Ser 186 2 0 1 (T) 0.005 12 rs17265551 c.1677 + 56C > T Intron 162 27 0 0.604 (T) 0.072 13 rs35604 c.1678-37G > A Intron 2 45 142 0.745 (G) 0.130 13 rs483352863* c.1678-34G > A Intron 188 1 0 1 (A) 0.003 13 rs35605 c.1684 T > C p.Leu562= 2 45 142 0.745 (T) 0.130 13 rs8187858 c.1704C > T p.Tyr568= 157 31 1 1 (T) 0.088 14 rs112282109 c.1898G > A p.Arg633Gln 187 1 0 1 (A) 0.003 16 rs8187863 c.2001C > T p.Ser667= 187 1 0 1 (T) 0.003 16 rs45511401 c.2012G > T p.Gly671Val 161 25 2 0.296 (T) 0.077 17 rs4148356 c.2168G > A p.Arg723Gln 181 9 0 1 (A) 0.024 19 rs45607032 c.2461-39_2461-38delAT Intron 179 9 0 1 (delAT) 0.024 19 rs2074087 c.2461-30C > G Intron 0 44 144 0.083 (C) 0.117 19 rs45492500 c.2461-27G > A Intron 172 14 2 0.056 (A) 0.048 21 rs11075296 c.2871 + 26C > T Intron 0 0 189 1 - 22 rs768191257 c.2876A > G p.Lys959Arg 187 1 0 1 (G) 0.003 22 rs3851716 c.3079 + 10G > A Intron 0 0 188 1 - 22 rs34794353 c.3079 + 24C > T Intron 187 1 0 1 (T) 0.003 22 rs3887893 c.3079 + 62 T > C Intron 67 96 25 0.358 (C) 0.388 23 rs191017838 c.3171G > A p.Leu1057= 187 2 0 1 (A) 0.005 23 rs199773531 c.3196C > T p.Arg1066Trp 188 1 0 1 (T) 0.003 25 rs41278168 c.3591-5C > T Intron 187 1 0 1 (T) 0.003 27 rs200922662 c.3886C > T p.Arg1296Trp 187 1 0 1 (T) 0.003 27 rs201533167 c.3901C > T p.Arg1301Cys 187 1 0 1 (T) 0.003 Linkage disequilibrium analysis Based on full genotype sets of 44 polymorphic variants confirmed by Hardy-Weinberg equilibrium exact test (Table 2), linkage disequilibrium analysis using r2 and |D`| statistics was performed (Additional file 4). Login to comment
154 Bold variants signifies the ones which were validated by genotyping results Table 3 Summary of ABCC1 selected SNPs genotyping by HRM and comparing them with scanning results dbSNP ID Variant residue NM_004996.3: Intron/amino acid residue NP_004987.2: Observed genotypesa (n) HWE exact test P-valueb MAFc (genotyping) MAFc (scanning) Chi-square test P-valued R/R R/V V/V rs41395947 c.128G > C p.Cys43Se 380 0 0 1 - - - rs2230669 c.816G > A p.Pro272= 362 18 0 1 (A) 0.024 (A) 0.043 0.079 rs246221 c.825 T > C p.Val275 197 160 23 0.243 (C) 0.271 (C) 0.309 0.187 rs8187852 c.1057G > A p.Val353Met 379 0 0 1 - - - rs35587 c.1062 T > C p.Asn354= 204 142 33 0.247 (C) 0.274 (C) 0.332 0.044 rs35588 c.1218 + 8A > G Intron 190 160 30 0.709 (G) 0.289 (G) 0.325 0.214 rs60782127 c.1299G > T p.Arg433Ser 373 6 0 1 (T) 0.008 (T) 0.005 0.623 rs35605 c.1684 T > C p.Leu562= 13 105 262 0.588 (T) 0.172 (T) 0.130 0.063 rs8187858 c.1704C > T p.Tyr568= 325 55 0 0.242 (T) 0.072 (T) 0.087 0.374 rs45511401 c.2012G > T p.Gly671Val 346 28 3 0.007 (T) 0.045 (T) 0.077 0.038 rs4148356 c.2168G > A p.Arg723Gln 360 19 0 1 (A) 0.025 (A) 0.024 0.888 rs45517537 c.2581G > A p.Ala861Thr 380 0 0 1 - - - rs35529209 c.2965G > A p.Ala989Thr 378 0 0 1 - - - rs13337489 c.3140G > C p.Cys1047Ser 380 0 0 1 - - - rs28706727 c.3436G > A p.Val1146Ile 380 0 0 1 - - - rs2230671 c.4002G > A p.Ser1334= 204 140 32 0.296 (A) 0.271 (A) 0.277 0.850 rs28364006 c.4009A > G p.Thr1337Ala 380 0 0 1 - - - a Number of genotypes detected during this study, R - reference allele, V - variant allele. b P-value is consistent with Hardy-Weinberg equilibrium if P > 0.001. c Minor allele shown in brackets with its frequency. Login to comment