ABCMdb

ABCC7 p.His1085Arg

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II (maturation defect) <a href="https://www.ncbi.nlm.nih.gov/pubmed/26823392">Veit et al.</a>
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Publications

PMID: 10517260 [PubMed] Yoshimura K et al: "A Japanese patient homozygous for the H1085R mutation in the CFTR gene presents with a severe form of cystic fibrosis."

No. Sentence Comment

1 All rights reser6ed Letter to the Editor A Japanese patient homozygous for the H1085R mutation in the CFTR gene presents with a severe form of cystic fibrosis To the Editor: Cystic fibrosis (CF), the most common autosomal recessive disorder in Caucasians, has long been believed to be extremely rare in Orientals (1, 2). Login to comment
4 Here we describe a 15-year-old female Japanese patient who exhibited typical manifestations of CF and proved to be a homozygote for a very rare missense mutation H1085R. Login to comment
15 Direct sequencing of the band clearly demonstrated an alteration of the nucleotide residue at 3386 from A to G in a homozygous fashion, resulting in His to Arg change at the amino acid 1085, a missense mutation H1085R. Login to comment
17 Further evaluation for three polymorphic loci proved that the H1085R mutation allele was associated with a haplotype of (GATT)6 in intron 6a (8), (TG)12T7 in the TG repeat and polythymidine tract at the splice acceptor site in intron 8 (9, 10), and M470 (1540A) in exon 10 (11, 12), respectively. Login to comment
18 The mutation H1085R, first described by Mercier et al. Login to comment
20 However, no other CF patients with H1085R have been reported since then, implying that it is an extremely rare mutation (14). Login to comment
21 Consistent with the previous report that other mutations located in exon 17b, such as R1066L and M1101R, were usually associated with pancreatic insufficiency, the case presented here and the French case had pancreatic insufficiency, suggesting that the H1085R is also a severe allele [(13), personal communication]. Login to comment
23 One important point is that the H1085R alleles of these 2 patients are most likely to be recurrent Letter to the Editor Fig. 1. Login to comment
24 Screening for the CFTR mutation H1085R in the patient and her family. Login to comment
27 The positions of the RsaI restriction site in the normal CFTR allele are indicated by hatched arrows, whereas the new RsaI site generated by H1085R mutation is by a solid arrow. Login to comment
36 However, it remains to be further elucidated whether this haplotype is restricted to the H1085R mutation or widespread in any other CFTR genotypes in CF patients in Japan. Login to comment

PMID: 11504857 [PubMed] Chen JM et al: "A combined analysis of the cystic fibrosis transmembrane conductance regulator: implications for structure and disease models."

No. Sentence Comment

571 This stringent sequence conservation, coupled with a cluster of missense mutations identified in this subdomain (among which two homozygosities, H1085R [Yoshimura et al. 1999] and R1066C [Casals et al. 1997], cause a severe CF phenotype), undoubtedly underlies its functional significance within CFTR. Login to comment
947 A Japanese patient homozygous for the H1085R mutation in the CFTR gene presents with a severe form of cystic fibrosis. Login to comment

PMID: 11933191 [PubMed] Ravnik-Glavac M et al: "DHPLC screening of cystic fibrosis gene mutations."

No. Sentence Comment

42 The following mutations have been studied: exon 3: W57G, R74W, R75Q, G85E, 394delTT, 405+ 1G>A; exon 4: E92X, P99L, 441delA, 444delA, 457TAT>G, D110H, R117C, R117H, A120T, 541delC, 544delCA, Q151X, 621+1G>T, 662- 2A>C; exon 7: 1078delT, F331L, R334W, I336K, R347C, R347P, A349V, R352Q, 1221delCT; exon 10: S492F, Q493X, 1609delCA, deltaI507, deltaF508; exon 11: G542X, S549N, G551D, R553X, A559T, R560K, R560T; exon 13: K716X, Q685X, G628R, L719X; exon 17b: H1054D, G1061R, 3320ins5, R1066H, R1066L, R1070Q, 3359delCT, L1077P, H1085R, Y1092X; exon 19: R1162X, 3659delC, 3662delA, 3667del4, 3737delA, I1234V, S1235R, 3849G>A; exon 20: 3860ins31,S1255X,3898insC,3905insT,D1270N, W1282X, Q1291R; and exon 21: N1303H, N1303K, W1316X. Login to comment

PMID: 12124743 [PubMed] Salvatore F et al: "Genotype-phenotype correlation in cystic fibrosis: the role of modifier genes."

No. Sentence Comment

46 A series of mutations usually associated with pancreatic sufficiency have been identified and defined as ''mild`` with reference to pancreatic status [Kerem et al., 1989c]: G85E, G91R, R117H, E193K, P205S, R334W, T338I, R347H, R347L, R347P, R352Q, A455E, S492F, S549N, P574H, D579G, 711 þ 5 G > A, C866Y, F1052V, H1054D, R1066H, R1068H, H1085R, D1152H, S1159P, S1251N, F1286S, G1349D, 2789 þ 5 G > A, and 3849 þ 10kb C > T [Dean et al., 1990; Cutting et al., 1990a; Cremonesi et al., 1992; Highsmith et al., 1994]. Login to comment

PMID: 15084988 [PubMed] Naruse S et al: "A finger sweat chloride test for the detection of a high-risk group of chronic pancreatitis."

No. Sentence Comment

51 The 9 CF-causing mutations (R75X, Q98R, M152R, R347H, L441P, L571S, D979A, H1085R, and T1086I) in Japa- nese20,25-28 were screened by SNP typing with Masscode System (Shimadzu, Kyoto, Japan). Login to comment

PMID: 15121783 [PubMed] Fujiki K et al: "Genetic evidence for CFTR dysfunction in Japanese: background for chronic pancreatitis."

No. Sentence Comment

219 The nine CF causing (R75X, Q98R, M152R, R347H, L441P, L571S, D979A, H1085R, and T1086I) and two non-CF causing (Q1352H and R1453W) mutations in Japanese6 22-24 were screened by SNP typing with a Masscode system (Shimadzu, Kyoto, Japan) and confirmed by sequence analysis in positive and equivocal cases. Login to comment

PMID: 15141088 [PubMed] Cormet-Boyaka E et al: "Rescuing cystic fibrosis transmembrane conductance regulator (CFTR)-processing mutants by transcomplementation."

No. Sentence Comment

5 This transcomplementation effect required a specific match between the region flanking the disease-causing mutation and the complementing fragment; e.g., amino fragments complemented ⌬F508-CFTR but not H1085R (a carboxy- processing mutant), whereas a carboxy fragment complemented H1085R but not ⌬F508-CFTR. Login to comment
146 All results are representative of 3-10 experiments. by comparing the transcomplementation of the ⌬F508-CFTR mutant to that of a severe processing mutant (H1085R-CFTR) for which the mutation resides in the carboxy-terminal half of the protein (27). Login to comment
147 Unlike for ⌬F508-CFTR, the processing of H1085R-CFTR could not be rescued by coexpression with the 1-633 amino fragment (Figs. 4 D and F). Login to comment
148 Importantly, however, H1085R-CFTR could be transcomplemented by a carboxy-terminal fragment (837-1,480), which had no effect on the processing of ⌬F508-CFTR or the N-tail processing mutants (Figs. 4 D, E, and F). Login to comment
170 (D) ⌬F508-CFTR was not transcomplemented by a carboxy fragment (837-1,480), whereas H1085R-CFTR, a cytosolic-loop-4-processing mutant that associates with severe disease, was complemented by fragment 837-1,480 but not by fragment 1-633. Login to comment
193 We thank M. J. Welsh for the H1085R-CFTR construct. Login to comment

PMID: 15357566 [PubMed] Ngukam A et al: "A novel missense mutation A1081P in the cystic fibrosis transmembrane conductance regulator (CFTR) gene identified in a Laotian patient with congenital bilateral absence of the vas deferens."

No. Sentence Comment

4 Only a few CFTR mutations have been identified in that population (L88X, M152R, K166E, F508del, 1742delAC, 1525-18G>A, 1540del10, L568X, 1898ϩ1G>T, 1898ϩ5G>T, G970D, 451-458del8, 3121-2A>G, H1085R).1-6 We report here a novel missense mutation in a Laotian patient with congenital bilateral absence of the vas deferens (CBAVD). Login to comment

PMID: 15880796 [PubMed] Kerem E et al: "Pharmacological induction of CFTR function in patients with cystic fibrosis: mutation-specific therapy."

No. Sentence Comment

58 C-D565G II DF508 D1507 S549R S549I S549N S549R S945D S945L H1054D G1061R L1065P R1066C R1066M L1077P H1085R N1303K G85E III G551D S492F V520F R553G R560T R560S Y569D IV R117H, R117C, R117P, R117L D1152H, L88S, G91R, E92K, Q98R, P205S, L206W, L227R, F311L, G314E, R334W, R334Q, I336K, T338I, L346P, R347C, R347H, R347L, R347P, L927P, R1070W, R1070Q V 3849 þ 10 kb C ! Login to comment

PMID: 16417523 [PubMed] Loo TW et al: "The chemical chaperone CFcor-325 repairs folding defects in the transmembrane domains of CFTR-processing mutants."

No. Sentence Comment

4 To test if CFcor-325 promoted correct folding of the TMDs (TM domains), we selected two of the CL4 mutants (Q1071P and H1085R) for disulphide cross-linking analysis. Login to comment
5 Pairs of cysteine residues that were cross-linked in mature wild-type CFTR were introduced into mutants Q1071P and H1085R. Login to comment
6 The cross-linking patterns of the Q1071P or H1085R double cysteine mutants rescued with CFcor-325 were similar to those observed with mature wild-type double cysteine proteins. Login to comment
21 EXPERIMENTAL Construction and expression of mutants The cDNAs of wild-type and CL4 mutants (H1054D, G1061R, L1065P, R1066H, Q1071P, L1077P, H1085R and W1098R) were inserted into pcDNA3 vector (Invitrogen, Oakville, ON, Canada) as described previously [2]. Login to comment
23 Site-directed mutagenesis was used to introduce the Q1071P or H1085R mutations into the double cysteine mutants as described previously [11]. Login to comment
31 Disulphide cross-linking analysis The cDNAs of double cysteine mutants M348C(TM6)/T1142C- (TM12), T351C(TM6)/T1142C(TM12) and W356C(TM6)/ W1145C(TM12) constructed in wild-type, mutant Q1071P or mutant H1085R backgrounds were expressed in HEK-293 cells in the presence or absence of 3 µM CFcor-325. Login to comment
37 Briefly, cells expressing wild-type, mutant Q1071P or H1085R CFTR were grown with or without 3 µM CFcor-325. Login to comment
51 Accordingly, HEK-293 cells were transfected with mutants H1054D, G1061R, L1065P, R1066H, Q1071P, L1077P, H1085R or W1098R cDNAs. Login to comment
57 Expression of mutants H1054D, G1061R, R1066H, Q1071P, L1077P, H1085R and W1098R in the presence of 3 µM CFcor-325, however, induced expression of the 190 kDa mature protein. Login to comment
59 The chemical chaperone most efficiently rescued mutants R1066H, Q1071P, H1085R and W1098R as the amount of mature CFTR was approx. Login to comment
73 Two CL4 mutants, Q1071P and H1085R, were selected for cross-linking analysis because they were efficiently rescued with CFcor-325. Login to comment
74 Expression of mutants Q1071P or H1085R in the presence of CFcor-325 increased the amount of mature CFTR from less than 5% (in the absence of corrector) of total to more than 30% (Figures 2A and 2B). Login to comment
77 Mature 190 kDa CFTR, however, contains complex carbohydrate groups that are resistant to endo H but not to PNGase F. Accordingly, cells expressing wild-type, mutant Q1071P or mutant H1085R CFTR and grown in the presence or absence of CFcor-325 were extracted with SDS and then treated with endoglycosidases. Login to comment
78 Figure 3 shows that the 170 kDa immature CFTRs but not the 190 kDa mature CFTRs of wild-type, mutant Q1071P or mutant H1085R were sensitive to digestion with endo H. Login to comment
79 Both the 190 kDa mature and 170 kDa immature CFTRs were sensitive to digestion with PNGase F. Therefore expression of mutant Q1071P and H1085R in the presence of CFcor-325 promoted maturation and trafficking of the proteins from the ER to the Golgi where complex carbohydrates are added to the protein. Login to comment
81 Cells expressing wild-type, mutant Q1071P or mutant H1085R CFTRs were grown in the presence or absence of 3 µM CFcor-325. Login to comment
82 The cells Figure 3 Endoglycosidase digestion Whole cell SDS extracts of HEK-293 cells expressing wild-type, mutant Q1071P or mutant H1085R in the absence (-) or presence (+) of 3 µM CFcor-325 were treated with endo H (H), PNGase F (F) or no endoglycosidase (-). Login to comment
85 Figure 4 Cell surface labelling of CFTR (A) BHK cells (Control) or BHK cells stably expressing wild-type (Wild-type), mutant Q1071P or mutant H1085R CFTRs were grown in the presence (+) or absence (-) of 3 µM CFcor-325. Login to comment
93 Figure 4(A) Figure 5 Measurement of cAMP-stimulated iodide efflux activities Iodide efflux assays were performed on BHK cells stably expressing wild-type, mutant Q1071P or mutant H1085R CFTRs that were grown in the presence (+CFcor-325) or absence (untreated) of CFcor-325 as described in the Experimental section. Login to comment
94 The activity of mutant H1085R grown without CFcor-325 was similar to that of mutant Q1071P (untreated) and is omitted for clarity. Login to comment
97 Similarly, labelled mature 190 kDa CFTR proteins were detected in mutants Q1071P and H1085R only after rescue with CFcor-325. Login to comment
98 Figure 4(B) shows that there was good correlation between the amount of biotinylated CFTR proteins in mutants Q1071P and H1085R with the amount of mature CFTRs detected in Figure 2(B) (63 and 50% of wild-type respectively). Login to comment
99 Next, we tested whether mutant Q1071P or H1085R was active at the cell surface after rescue with CFcor-325. Login to comment
100 Mutants Q1071P and H1085R CFTRs were stably expressed in BHK cells because adherent cells are essential for the iodide efflux assays. Login to comment
102 The BHK cells stably expressing mutants Q1071P or H1085R were treated for 48 h with or without 3 µM CFcor-325 and then used in iodide efflux assays. Login to comment
106 In contrast, BHK cells stably expressing CFTR mutants Q1071P or H1085R demonstrated iodide efflux activity only when grown in the presence of CFcor-325 (Figure 5). Login to comment
109 Accordingly, M348C(TM6)/T1142C(TM12), T351C(TM6)/ T1142C(TM12) or W356C(TM6)/W1145C(TM12) mutations were introduced into a Q1071P or H1085R CFTR background. Login to comment
110 Mutants Q1071P or H1085R containing M348C(TM6)/T1142C- (TM12), T351C(TM6)/T1142C(TM12) or W356C(TM6)/ W1145C(TM12) mutations were then transiently expressed in HEK-293 cells in the presence or absence of 3 µM CFcor-325 for 48 h and then treated with the homobifunctional cross-linkers M5M, M8M or M17M. Login to comment
115 Mature mutant Q1071P/ M348C(TM6)/T1142C(TM12) protein was cross-linked with Figure 6 Disulphide cross-linking analysis of CFTR processing mutants HEK-293 cells expressing mutants Q1071P/M348C(TM6)/T1142C(TM12), Q1071P/T351C- (TM6)/T1142C(TM12) and Q1071P/W356C(TM6)/W1145C(TM12) (A), mutants H1085R/ M348C(TM6)/T1142C(TM12), H1085R/T351C(TM6)/T1142C(TM12) and H1085R/W356C- (TM6)/W1145C(TM12) (B) or wild-type, mutant Q1071P or mutant H1085R (C) were incubated for 48 h with (+) or without (-) 3 µM CFcor-325. Login to comment
120 Similar results were observed with the H1085R double cysteine mutants (Figure 6B). Login to comment
122 In contrast, wild-type, mature mutant Q1071P or H1085R CFTRs lacking the introduced cysteine residues was not cross-linked by M5M, M8M or M17M (Figure 6C). Login to comment
129 Similar results were observed with mutant H1085R/M348C(TM6)/T1142(TM12) (results not shown). Login to comment

PMID: 16624886 [PubMed] Wang Y et al: "Specific rescue of cystic fibrosis transmembrane conductance regulator processing mutants using pharmacological chaperones."

No. Sentence Comment

30 Wild-type, ⌬F508, and H1085R CFTR cDNAs were inserted into the pcDNA3 vector (Invitrogen Canada Inc., Burlington, ON, Canada) as described previously (Loo et al., 2006). Login to comment
138 It has been reported that it was possible to specifically rescue CFTR processing mutants containing a processing mutation in the front half of the protein (⌬F508) by coexpression with a CFTR NH2-half molecule whereas processing mutations located in the back half of the protein (H1085R) could be rescued by coexpression with a CFTR COOH-half molecule (Cormet-Boyaka et al., 2004). Login to comment
140 Therefore, we tested whether a specific CFTR pharmacological chaperone such as CFpot-532 could rescue a CFTR processing mutant with a mutation in the COOH half of the molecule (H1085R). Login to comment
141 HEK 293 cells transiently expressing CFTR processing mutant H1085R were incubated in the presence or absence of 10 ␮M CFpot-532 for 48 h. Whole-cell extracts were subjected to immunoblot analysis. Login to comment
142 Figure 7 shows that expression of CFTR H1085R in the presence of 10 ␮M CFpot-532 promoted maturation of the protein. Login to comment
170 HEK 293 cells were transiently transfected with CFTR wild-type or H1085R cDNAs and then expressed in the absence (-) or presence (ϩ) of 10 ␮M CFpot-532 for 48 h. Whole-cell extracts were subjected to immunoblot analysis with a CFTR polyclonal antibody. Login to comment

PMID: 17535157 [PubMed] Wang Y et al: "Additive effect of multiple pharmacological chaperones on maturation of CFTR processing mutants."

No. Sentence Comment

33 MATERIALS AND METHODS Construction and expression of mutants The construction of F508 and H1085R CFTR cDNAs and insertion into the pcDNA3 vector (Invitrogen Canada, Burlington, ON, Canada) was described previously [23]. Login to comment
34 The CFTR cDNAs coding for NBD2 (where NBD2 is nucleotide-binding domain 2), H1085R CFTR (residues 1-1196) as well as P-gp cDNAs for mutant P709G P-gp and P-gp truncation mutants NBD2 P-gp (residues 1-1023), TMD1+2 {TMD1 [N-terminal TMD (TM domain) containing TM segments 1-6] and TMD2 (C-terminal TMD containing TM segments 7-12)}, P-gp (residues 1-379 plus 681-1025) and TMD1 P-gp (residues 1-379) were modified to contain the epitope for monoclonal antibody A52 at the C-terminal ends and subcloned into the mammalian expression vector pMT21 as described previously [21]. Login to comment
103 Figure 4 Effect of correctors on maturation of CFTR processing mutant H1085R CFTR processing mutant H1085R was expressed for 24 h in the absence (-) or presence (+) of 3 µM VRT-325, 10 µM corr-2b, 10 µM corr-4a, combination of 3 µM VRT-325 and 10 µM corr-2b, combination of 3 µM VRT-325 and 10 µM corr-4a, or combination of 10 µM corr-2b and 10 µM corr-4a respectively. Login to comment
112 First, we tested the correctors on a CFTR processing mutant (H1085R) with a mutation in the fourth cytoplasmic loop (CL4) connecting TM segments 10 and 11 in the CO2H half of the molecule, where a relatively large number of clinically relevant processing mutations are found [35]. Login to comment
113 The H1085R CFTR mutant was selected for use in our initial studies because expression of its cDNA in HEK-293 cells yields a small amount of mature protein (approx. Login to comment
115 Therefore the use of mutant H1085R in the presence of combinations of correctors would enable us to determine if the compounds increase or decrease the efficiency of maturation. Login to comment
116 CFTR processing mutant H1085R was transiently expressed in HEK-293 cells and incubated in the presence or absence of 3 µM VRT-325, 10 µM corr-2b and 10 µM corr-4a, combination of VRT-325 and corr-2b, combination of VRT-325 and corr-4a, or combination of corr-2b and corr-4a for 24 h. Whole cell extracts were subjected to immunoblot analysis. Login to comment
119 Expression of mutant H1085R CFTR in the presence of a combination of VRT-325 and corr-2b caused the largest increase in maturation efficiency as 48% of total CFTR was present as mature protein. Login to comment
122 It has been postulated that correctors promote maturation of CFTR processing mutants by promoting interactions between the Figure 5 Effect of correctors on maturation of CFTR truncation mutant NBD2 H1085R HEK-293 cells were transfected with NBD2 H1085R followed by a 24 h incubation in the absence (-) or presence (+) of 3 µM VRT-325, 10 µM corr-2b, 10 µM corr-4a, combination of 3 µM VRT-325 and 10 µM corr-2b, or combination of 3 µM VRT-325 and 10 µM corr-4a respectively. Login to comment
126 To determine if interactions between the NBDs were required for the enhanced maturation of mutant H1085R in the presence of VRT-325 together with corr-2b or corr-4a, we utilized an H1085R mutant that lacked NBD2 ( NDB2 H1085R CFTR) (Figure 1B). Login to comment
127 The NDB2 H1085R CFTR mutant was transfected into HEK-293 cells and expressed in the presence of VRT-325, corr-2b, corr-4a, VRT-325 plus corr-2b, VRT-325 plus corr-4a or no correctors for 24 h. Whole cell extracts were then subjected to immunoblot analysis. Login to comment

PMID: 17680777 [PubMed] Skach WR et al: "Pharmacological chaperoning: two 'hits' are better than one."

No. Sentence Comment

47 Steady-state levels (determined by immunoblotting) of the CFTR mutant H1085R revealed that combinations of corr-2b/VRT325 or corr-4a/ corr-2b rescued more CFTR than each compound independently. Login to comment
48 Similar additive effects were observed for combinations of corr-2b/VRT325 or corr-4a/corr-2b for the CFTR mutant H1085R, in which the second nucleotide-binding domain was removed. Login to comment

PMID: 18193900 [PubMed] Cheung JC et al: "Misfolding of the cystic fibrosis transmembrane conductance regulator and disease."

No. Sentence Comment

90 In some additional examples, a number of mutations found in the fourth intracellular loop (H1054D, G1061R, L1065P, R1066C/H/L, Q1071P, L1077P, H1085R, W1098R, M1101K/ R) also affect the biosynthetic processing of CFTR (although function was not tested) (73); some intracellular loop 4 mutants (F1052V, K1060T, A1067T, G1069R, R1070Q/W) can process CFTR to the complex-glycosylated ("Band C") form but have altered channel activity compared to wild type. Login to comment

PMID: 19620404 [PubMed] Cormet-Boyaka E et al: "A truncated CFTR protein rescues endogenous DeltaF508-CFTR and corrects chloride transport in mice."

No. Sentence Comment

215 As previously reported, the rescue was specific to mutations located in the amino-terminal part of CFTR (N-tail and NBD1), as the CFTR fragment 1-633 did not rescue a CFTR mutant harboring a mutation located close to the carboxyl tail of CFTR, H1085R-CFTR (14). Login to comment

PMID: 19652440 [PubMed] Izumikawa K et al: "Unique mutations of the cystic fibrosis transmembrane conductance regulator gene of three cases of cystic fibrosis in Nagasaki, Japan."

No. Sentence Comment

16 The other novel or rare mutations such as R347H, D 979A, 1724delAG, H1085R, M152R and 1540del10 have The Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Department of Respiratory Medicine, National Hospital Organization Minami-Kyushu National Hospital, Kagoshima, Department of Respiratory Medicine, National Hospital Organization Ureshino Medical Center, Ureshino, Department of Laboratory Medicine, Nagasaki University School of Medicine, Nagasaki and Department of Respiratory Medicine, Respiratory Center, Toranomon Hospital, Tokyo Received for publication January 20, 2009; Accepted for publication April 14, 2009 Correspondence to Dr. Koichi Izumikawa, koizumik@nagasaki-u.ac.jp Figure1. Login to comment

PMID: 20110398 [PubMed] Bertrand J et al: "Identification of a novel water-soluble activator of wild-type and F508del CFTR: GPact-11a."

No. Sentence Comment

45 Human lung tissues were obtained from seven patients: three non-CF males with a mean age of 61 yrs, a non-CF female aged 65 yrs, a F508del/F508del-CFTR male aged 37 yrs, a F508del/R1066C female aged 26 yrs and a F508del/ H1085R-CFTR female aged 23 yrs. Login to comment
118 The mutation H1085R is a severe and rare missense mutation identified by MERCIER et al. [32]. Login to comment
119 H1085R-CFTR protein presents a trafficking defect which can be corrected by F508del-CFTR corrector [33]. Login to comment
155 Structural determinants for CFTR activation In the chemical structure of GPact-11a (fig. 1b), three main elements can be distinguished: 1) the purine aromatic heterocycles composed of fused pyrimidine and imidazole rings; -15 ΔFx 35 25 15 5 -5 45 CF-KM4 F508del-CFTR HEK-293 CFPACCuFi-1CF15 -5 ΔFx 15 10 5 0 20 F508del/ H1085R F508del/ F508del F508del/ R1066C 0 Activation% 80 60 40 20 100 -2-3-4-5 Log [GPact-11a] M -6-7 Fx 40 20 0 -20 60a) b) c) d) 2015105 Time min GPact-11a CFTRInh-172 0 ● ● ● ● ● ● ● ● ●●●●●●●●●●●●● ●● ●●● ●●●● ● ● ● ● ● ● ●● ● ● ● ● ● FIGURE 5. Login to comment
161 d) Effect of GPact-11a on human bronchial ciliated epithelial cells cells freshly isolated from lungs with different cystic fibrosis genotypes (F508del/F508del-CFTR (n54), F508del/R1066C (n57) and F508del/H1085R-CFTR (n59)). Login to comment

PMID: 21520952 [PubMed] Loo TW et al: "Benzbromarone stabilizes DeltaF508 CFTR at the cell surface."

No. Sentence Comment

22 The H1085R mutation is located in TMD2 within the intracellular loop (ICL) connecting TM10 and TM11.24 Mature protein was observed when H1085R CFTR was expressed in the presence of 38À100 μM benzbromarone (Figure 1A). Login to comment
26 To test if the CFTR mutants were active after benzbromarone rescue, we performed iodide efflux assays on BHK cells stably expressing ΔF508, H1085R, or wild-type CFTR proteins. Login to comment
27 It was found that both ΔF508 and H1085R exhibited forskolin-activated iodide efflux after rescue with benzbromarone (Figure 1B). Login to comment
47 (A) Immunoblot analysis of cells expressing CFTR mutant ΔF508 or H1085R or the P-gp G251V processing mutant after treatment with the indicated concentrations of benzbromarone (Benz) for 40 h. Login to comment
48 (B) Iodide efflux assays performed on BHK cells stably expressing wild-type, ΔF508, or H1085R CFTR. Login to comment

PMID: 21594776 [PubMed] Loo TW et al: "Repair of CFTR folding defects with correctors that function as pharmacological chaperones."

No. Sentence Comment

147 HEK 293 cells expressing the CFTR processing mutants, F508del, Q1071P, or H1085R, were incubated with media containing no corrector (-) or 0.05 mM VX-325 plus 0.015 mM corr-4a (+). Login to comment

PMID: 17331079 [PubMed] Alonso MJ et al: "Spectrum of mutations in the CFTR gene in cystic fibrosis patients of Spanish ancestry."

No. Sentence Comment

52 Mutation 0.46-0.35 9 c.1078delT #, p.R347P # 8 p.G85V, c.621 + 1G > T #, p.S549R (T > G) #, p.R553X #, c.3849 + 10kbC > T # 7 p.R347H #, c.1812-1G > A, p.R709X 0.30-0.10 6 p.H199Y, p.P205S, 5 p.R117H #, p.G551D #, p.W1089X, p.Y1092X, CFTR50kbdel 4 c.296 + 3insT, c.1717-1G > A #, c.1949del84, c.3849 + 1G > A 3 p.E92K, c.936delTA, c.1717-8G > A, c.1341G > A, p.A561E, c.2603delT, p.G1244E, [p.D1270N; p.R74W] 2 p.Q2X, p.P5L, CFTRdele2,3, p.S50P, p.E60K, c.405 + 1G > A, c.1677delTA, p.L558S, p.G673X, p.R851X, p.Y1014C, p.Q1100P, p.M1101K, p.D1152H, CFTRdele19, p.G1244V, p.Q1281X, p.Y1381X <0,1 1 c.124del23bp, p.Q30X, p.W57X, c.406-1G > A, p.Q98R, p.E115del, c.519delT, p.L159S, c.711 + 3A > T, p.W202X, c.875 + 1G > A, p.E278del, p.W361R, c.1215delG, p.L365P, p.A399D, c.1548delG, p.K536X, p.R560G, c.1782delA, p.L571S, [p.G576A; p.R668C], p.T582R, p.E585X, c.1898 + 1G > A, c.1898 + 3A > G, c.2051delTT, p.E692X, p.R851L, c.2711delT, c.2751 + 3A > G, c.2752-26A > G, p.D924N, p.S945L, c.3121-1G > A, p.V1008D, p.L1065R, [p.R1070W; p.R668C], [p.F1074L; 5T], p.H1085R, p.R1158X, c.3659delC #, c.3667del4, c.3737delA, c.3860ins31, c.3905insT #, c.4005 + 1G > A, p.T1299I, p.E1308X, p.Q1313X, c.4095 + 2T > A, rearrangements study (n = 4) Mutations identified in CF families with mixed European origin: c.182delT, p.L1254X, c.4010del4. Login to comment

PMID: 22138447 [PubMed] Loo TW et al: "Corrector-mediated rescue of misprocessed CFTR mutants can be reduced by the P-glycoprotein drug pump."

No. Sentence Comment

14 Here, we tested our predictions that (1) other CFTR mutants such V232D and H1085R were more stable at the cell surface than DF508 CFTR after low temperature rescue and (2) the advantages of rescue with specific correctors (pharmacological chaperones) are that they may stabilize DF508 CFTR and increase the effectiveness of the correctors by bypassing drug pumps such as P-glycoprotein (P-gp) (increased bioavailability). Login to comment
15 It was found that the stability of mutants V232D and H1085R after low-temperature (30 8C) rescue was about 10-fold higher than DF508 CFTR. Login to comment
37 We show that rescue of DF508 CFTR with specific correctors resulted in a more stable protein and mutations such as V232D and H1085R are different from DF508 because they do not destabilize the rescued form of mature CFTR. Login to comment
67 To test the effect of P-gp on rescue of CFTR, cells were cotransfected with the H1085R CFTR processing mutant plus wild-type P-gp or an inactive P-gp mutant containing mutations to the catalytic carboxylate residues (E556Q/E1201Q) [26]. Login to comment
106 (B) Samples of cells expressing DF508, V232D or H1085R CFTR mutants or P-gp mutant G268V and grown in the absence (À) or presence (+) of 20 mM VX-809 for 18 h were subjected to immunoblot analysis. Login to comment
120 We also tested whether correctors would promote maturation of CFTR mutants DF508, V232D and H1085R in HEK 293 cells in parallel because it has been reported that CFTR rescue depends on the cell system used [38-41]. Login to comment
121 Mutants V232D (TMD1) and H1085R (TMD2) were included because they are processing mutations located in different domains of CFTR (Fig. 1A) and both yield active proteins after rescue [16,42,43]. Login to comment
181 Degradation of wild-type, V232D, and H1085R CFTRs was monitored over time at 37 8C. Login to comment
185 Fig. 8. Effect of wild-type P-gp expression (+P-gp) on rescue of CFTR mutant H1085R with corrector KM11060. Login to comment
203 To test if other CF mutations affect stability of CFTR, mutants V232D (TMD1) and H1085R (TMD2) were selected for study as both are processing mutations that yield active proteins after rescue [16,43]. Login to comment
204 Accordingly, we examined the stability of mutants V232D and H1085R after low-temperature rescue. Login to comment
205 Cells expressing wild-type, V232D, or H1085R CFTRs were expressed at low temperature to promote maturation of the protein. Protein synthesis was stopped by addition of cycloheximide, and turnover of the protein was monitored after incubation for 0-32 h at 37 8C (Fig. 7). Login to comment
206 It was observed that the half-lives of the mature forms of V232D and H1085R were at least 10-fold longer (about 14 and 12 h, respectively) than DF508 CFTR (about 1 h, Fig. 4). Login to comment
208 The V232D TMD1 and H1085R mutations may have less effect on the stability of mature CFTR because they have more localized effects on protein folding in the TMD1 and TMD2 domains, respectively. Login to comment
210 3.4. Effect of P-gp expression on rescue of CFTR mutant H1085R We predicted that the efficiency of CFTR rescue with correctors would be reduced if the correctors were also substrates of drug pumps such as P-gp. Login to comment
215 To test if P-gp expression would alter CFTR rescue with KM11060, mutant H1085R was expressed in the presence of various concentrations of KM11060 in the presence of an inactive P-gp containing mutations to the catalytic carboxylate residues (E556Q/E1201Q) in the nucleotide binding domains [26] or in the presence of wild-type P-gp. Login to comment
216 CFTR mutant H1085R was tested because ithas a higher efficiency of rescue with correctors compared to DF508 CFTR [16] and the mature protein is more stable (Fig. 7). Login to comment
219 The R50 for KM11060 when H1085R CFTR was expressed with noexogenous P-gp was similar to that obtained with inactive P-gp (data not shown) suggesting that expression of P-gp did not adversely affect the cellular folding machinery. Login to comment

PMID: 18556774 [PubMed] Schrijver I et al: "Multiplex ligation-dependent probe amplification identification of whole exon and single nucleotide deletions in the CFTR gene of Hispanic individuals with cystic fibrosis."

No. Sentence Comment

92 on.ca/cftr/) and is considered a specific African-American mutation.24 Finally, mutation H1085R has been described repeatedly (http://www.genet.sickkids.on.ca/cftr/) and was identified in one homozygous Japanese patient who was born to consanguineous parents.28 Thus, of the 22 mutations described in non-Caucasian populations, at least six (27.3%) are recurrent and may be specific to non-Caucasian populations. Login to comment
112 Mutations under MLPA Ligation Sites Exon Probe length (nt) Ligation site sequence Mutations in area of ligation site sequence* 1,5Ј UTR 154 5Ј-GAGCAAAT-TTGGGGCC-3Ј N/A 1,5Ј UTR 238 5Ј-AAAGGGTT-GAGCGGCA-3Ј 2 198 5Ј-TTGGTATA-TGTCTGAC-3Ј (5) 3 136 5Ј-CTGCTAGT-GTTGCCAA-3Ј (3) 3 220 5Ј-TTCAAAGA-AAAATCCT-3Ј 4 247 5Ј-AGAATCAT-AGCTTCCT-3Ј 444delA, African; 451del8, Chinese; (6) 5 346 5Ј-AAATAAGT-ATTGGACA-3Ј Q179K, Hispanic (7) 6a 274 5Ј-GAGTTGTT-ACAGGCGT-3Ј L218X, Pakistani (4) 6b 301 5Ј-ATTTTCAA-TCATTTCT-3Ј 935delA, Hispanic; 936delTA, Hispanic (3) 7 337 5Ј-ACTTCAAT-AGCTCAGC-3Ј S307N, Turkish (9) 8, IVS 8 364 5Ј-TTTCTAGA-TTAAGAAG-3Ј N/A 9, IVS 8 391 5Ј-TCCATCAC-ACTGGTAG-3Ј N/A 10 463 5Ј-TCCACTGT-GCTTAATT-3Ј H484Y, Hispanic; S485C, Chinese-Caucasian (5) 11 418 5Ј-CAGAGAAA-GACAATAT-3Ј K536X, Iranian; 1742delAC, Japanese (5) 12, IVS 12 292 5Ј-TGCATTTT-ACCTCTTG-3Ј N/A 13 142 5Ј-CAGATTCT-GAGCAGGG-3Ј (1) 14a 160 5Ј-GTATGTGT-TCCATGTA-3Ј (3) 14b 178 5Ј-CTGCTTCT-TTGGTTGT-3Ј 2766del8, Tunisian (1) 15 204 5Ј-GCTTGCTA-TGGGATTC-3Ј (1) 16, IVS 16 229 5Ј-GATGTAAT-AGCTGTCT-3Ј N/A 17a 256 5Ј-TGCAACAA-AGATGTAG-3Ј 3171delC, Hispanic; 3173delAC, Turkish; F1016S, Hispanic (5) 17b 283 5Ј-CAGTATGT-AAATTCAG-3Ј H1085R, Japanese (4) 18 310 5Ј-CCATGAAT-ATCATGAG-3Ј M1137R, Hispanic (6) 19 353 5Ј-TCTGTGTA-TTTTGCTG-3Ј 3791delC, African-American (2) 20 382 5Ј-CTTGGGAT-TCAATAAC-3Ј 3960delA, Hispanic (2) 21 409 5Ј-TGCAACTT-TCCATATT-3Ј W1316X, African-American (2) 22 436 5Ј-GAACAGTT-TCCTGGGA-3Ј No mutations 23 148 5Ј-CCAGCATT-GCTTCTAT-3Ј M1407T, Turkish; E1409K, Hispanic (2) 24 190 5Ј-ATCCAGAA-ACTGCTGA-3Ј No mutations 24 172 5Ј-CTCCTCTT-TCAGAGCA-3Ј UTR, untranslated region; IVS, intervening sequence; N/A, not applicable, probes not in coding region; No mutations, no reported mutations are present in the area of the ligation site sequence, regardless of ethnicity. Login to comment
128 In all, at least six (444delA, 935delA, 936delTA, 2766del8, 3791delC, and H1085R) of 22 mutations identified in non-Caucasians (27.3%) are recurrent and may be specific to non-Caucasian populations. Login to comment

PMID: 10923036 [PubMed] Claustres M et al: "Spectrum of CFTR mutations in cystic fibrosis and in congenital absence of the vas deferens in France."

No. Sentence Comment

107 f 306insA, W79X, R117C, P205S, L227R, I336K, 1248+1G>A, 1609delCA, 1717-8G>A, S549R(T>G), S549N, 1812-1G>A, P574H, 2176insC, R709X, E827X, D836Y, 3007delG, L1065P, L1077P, H1085R, M1101K, 4021insT. Login to comment

PMID: 8702904 [PubMed] Cotten JF et al: "Effect of cystic fibrosis-associated mutations in the fourth intracellular loop of cystic fibrosis transmembrane conductance regulator."

No. Sentence Comment

81 For example, levels of mature F1052V were similar to wild-type, whereas L1065P, R1070Q, and H1085R were similar to ⌬F508 in that they produced little mature protein (Fig. 2, A and B). Login to comment
222 For example, H1085R is misprocessed like ⌬F508, yet it is reported to occur in a patient with a pancreatic sufficient phenotype. Login to comment
80 For example, levels of mature F1052V were similar to wild-type, whereas L1065P, R1070Q, and H1085R were similar to DF508 in that they produced little mature protein (Fig. 2, A and B). Login to comment
221 For example, H1085R is misprocessed like DF508, yet it is reported to occur in a patient with a pancreatic sufficient phenotype. Login to comment

PMID: 8662892 [PubMed] Seibert FS et al: "Disease-associated mutations in the fourth cytoplasmic loop of cystic fibrosis transmembrane conductance regulator compromise biosynthetic processing and chloride channel activity."

No. Sentence Comment

130 D: छ, WT; E, Q1071P; छϩ, W1098R; Ⅺ, H1085R; Ç, M1101K; µ, M1101R; Q, control; É, L1077P. Login to comment
136 D: L, WT; E, Q1071P; L 1, W1098R; M, H1085R; &#c7;, M1101K; &#b5;, M1101R; Q, control; &#c9;, L1077P. Login to comment

PMID: 8844213 [PubMed] Morral N et al: "Haplotype analysis of 94 cystic fibrosis mutations with seven polymorphic CFTR DNA markers."

No. Sentence Comment

105 CFTR Haplotypes for Diallelic and Multiallelic DNA Markers for 94 CF Mutations" J44-GATT- 8CA-17BTA- No. of T854-TUB20 17BCA Mutation chromosomes % Normal Laboratory Reference 2-7-1-2 17-47-13 (55.4%) 17-46-13 17-45-13 17-34-13 17-32-13 17-31-14 17-31-13 17-29-14 17-28-13 16-48-13 16-46-14 16-46-13 16-45-13 16-44-13 16-35-13 16-33-13 16-32-13 16-31-14 16-31-13 16-30-13 16-29-13 16-26-13 16-25-13 16-24-13 14-31-13 1-7-2-1 17-7-17 (16.8%) R334W R334W 3860ins31 G1244E R1162X R1162X R1162X G91R MllOlK R347P R334W R117C E92K 3849+lOkbC+T 3293delA 1811+1.6kb A-tG 1811+1.6kb A-tG 2184insA P205S 3659delC G673X 11005R I336K W58S R347P W846X 405+1-A G178R 3905insT R1162X R347H 3100insA E60X 1078delT 4005+1-A K710X 1677delTA H199Y 3601-2AjG 3850-3T+G 3272-26A-tG 3850-1-A 1812-1-A R117H L1059X S492F Y1092X Y569H 3732delA C866Y 711+1G+T 711+1-T G85E 1949del84 2789+5-A H1085R W1282X R1066C 2043delG V456F 2 1 1 1 2 1 6 2 2 1 2 1 1 2 1 1 4 1 1 1 3 2 1 1 1 1 1 1 2 7 1 1 1 1 2 1 1 3 19 3 3 1 1 2 1 1 5 1 1 1 1 3 6 3 5 1 13 2 1 1 - 0.48 0.48 - - - 0.24 - - - 2.65 2.40 1.93 2.65 1.68 2.65 0.72 13.94 13.46 1.93 - 0.72 0.24 3.37 - b b fP fP fP t b,fb.fP h fb t h t h h fP fP b.h b h h b h h h h h fb fb,fP.t fP fP fP9t fP b t fPh b h fb b.fb,h fb*fP b,fP h h t h fb fb,fp,h.t fP fP fb t b.fP,t b,fb,h,t b f b h h fb b,fb.fP,h fP h h Gasparini et al. (1991b) Chilldn et al. (1993a) Devoto et al. (1991) Gasparini et al. (1991b) Dork et al. (1993a) Guillermit et al. (1993) Zielenski et al. (1993) Dean et al. (1990) Dork et al. (1994a) Nunes et al. (1993) Highsmith et al. (1994) Ghanem et al. (1994) Chilldn et al. (1995) Dork et al. (1994a) Dork et al. (1993a) Chilldn et al. (1993b) Kerem et al. (1990) Dork et al. (1994a) Dork et al. (1994a) Cuppenset al. (1993) Fanen et al. (1992) Maggio et al. (personal communication) Audrezet et al. (1993) Vidaud et al. (1990) Dork et al. (1993b) Zielenski et al. (1991a) Chilldn et al. (1994b) Malik et al. (personal communication) Cremonesi et at. Login to comment

PMID: 7543317 [PubMed] Pignatti PF et al: "Increased incidence of cystic fibrosis gene mutations in adults with disseminated bronchiectasis."

No. Sentence Comment

31 List of CFTR gene mutations and DNA polymorphisms screened Mutations R75Q/X/L, G85E, 394deITT 457TAT->G, R117H 621 + 1G->T 711 + 5G->A L206W 875 + 40 A->G 936 del TA 1001 + 11C->T R334W, R347 P/H/L, 1154insTC A455E, V456F DF5O8 1717-IG->A, 1717-8G->A G542X, G551D, Q552X, R553X P574H 1898 + 3A->G 2183 AA->G, 2184delA, R709X D836Y, 2694 T/G 2752-22 A/G 2789 + 5 G->A, 2790-2 A-»G Q890X 3041-71 G/C 3132delTG 3271 + 18 C-»T, 3272-26 A->G H1054D, G1061R, R1066C/H, A1067T, H1085R, Y1092X, 3320 ins5 D1152H R1162X, 3667ins4, 3737delA, 11234V 3849 + 10 kb C-»T, 3850-1 G-»A SI25IN, S1255P, 3905insT, 3898insC, D127ON, W1282X, R1283M, 4002 A/G 4005 + 1 G-»A N1303 K/H, 4029 A/G D1377H Q1411 X 4404 C/T, 4521 G/A Location e 3 e 4 i 4 i 5 e 6a i 6a e 6b i 6b e 7 e 9 e 10 i 10 e 11 e 12 i 12 e 13 e 14a i 14a i 14b e 15 i 15 e 17a i 17a e 17b e 18 e 19 i 19 e 20 i 20 e2l e 22 e 23 e24 Listing is in order of location along the CFTR gene, e = exon; i = intron. Login to comment

PMID: 7529319 [PubMed] Mercier B et al: "A cluster of cystic fibrosis mutations in exon 17b of the CFTR gene: a site for rare mutations."

No. Sentence Comment

19 Most of these are missense mutations and as no functional test has been 732 Table 1 Mutations identified in exon 17b of the CFTR gene Mutation Nucleotide Modificationl Ethnic Rcferencesposition ongini (No) 3271-1 G--A 3272-1 G-A Belgian (1) 11F1052V 3286 T-G Belgian (1) 11HI054D 3292 CG French (1) 13G1061R 3313 G-C French (1) 113320 Dup 3320 Duplication of Breton (1) 6 CTATG R1066C 3328 CT French (1) 14 R1066L R1066H A1067T G1069R R1070Q 3359 del CT L1077P H1085R W1089X Y1092X M1IOIR 3329 3329 3331 3337 3341 G-T G-+A G-A G,A G--A 3359 3362 3386 3398 3408 3434 del CT T--C A-.G G-+A C +A T--G Spanish (5) French (1) Breton (1) Breton (1) Bulgarian (1) Bulgarian (3) Rumanian (1) Albanian (1) French (1) Italian (1) French (1) Spanish (1) French (4) Turkish (1I) * Bozon et al, personal communication. Login to comment

PMID: 7525963 [PubMed] Chevalier-Porst F et al: "Mutation analysis in 600 French cystic fibrosis patients."

No. Sentence Comment

21 Among the 104 other CFTR mutations tested on the 373 non-AF508 CF chromosomes, none of the following 58 mutations were found: G91R, 435 insA, 444delA, D11OH, 556delA, 557delT, R297Q, 1154insTC, R347L, R352Q, Q359K/T360K, 1221delCT, G480C, Q493R, V520F, C524X, 1706dell7, S549R (A-C), S549N, S549I, G551S, 1784delG, Q552X, L558S, A559T, R560T, R560K, Y563N, P574H, 2307insA, 2522insC, 2556insAT, E827X, Q890X, Y913C, 2991de132 (Dork et al, personal communication), L967S, 3320ins5, 3359delCT, H1085R, R1158X, 3662delA, 3667del4, 3667ins4, 3732delA, 3737delA, W1204X, 3750delAG, I 1234V, Q1238X, 3850- 3T-+G, 3860ins31, S1255X, 3898insC, D1270N, R1283M, F1286S, 4005 + I G-A. Forty-six other mutations were found on at Distribution of CFTR mutations found in our sample ofpopulation (1200 CF chromosomes) Mutations tested No of CF chromosomes Haplotypes Method with the mutation XV2C-KM19 (% of total CF alleles) Exon 3: G85E 4 (033) 3C HinfI/ASO394delTT 2 2B PAGEExon 4: R117H 1 B ASOY122X 2 2C MseI/sequenceI148T 1 B ASO621+IG-J* 1 B MseIIASOExon 5: 711+1G--T 8(07) 8A ASOExon 7: AF311 1 C PAGE/sequencelO78delT 5 (0-42) 5C PAGE/ASOR334W 5 (0-42) 2A,2C,ID MspIlASOR347P 5 (042) 5A CfoI/NcoIR347H 1 Cfol/sequenceExon 9: A455E 1 B ASOExon 10: S492F I C DdeI/sequenceQ493X 1 D ASOl609deICA 1 C PAGE/Ddel/sequenceA1507 3 (025) 3D PAGE/ASOAF508 827 (69) 794B,30D,2C,IA PAGEl677delTA 1 A PAGE/sequenceExon I11: 1717-IG--.A 16(1-3) 14B Modified primers + AvaIIG542X 40 (3-3) 29B,5D,2A Modified primers + BstNiS549R(T--*G) 2 2B ASOG551D 3 (025) 3B HincII/Sau3AR553X 10(0-8) 6A,1B,2C,ID Hincll/sequenceExon 12: 1898+IG--A 1 C ASO1898+ IG-C 2 IC ASOExon 13: l9l8deIGC 1 A PAGE/sequence1949de184 I C PAGE/sequenceG628R(G-+A) 2 2A Sequence2118de14 I c PAGE/sequence2143de1T 1 B PAGE/modified primers2184de1A+2183A--*G 11 (0-9) lIB PAGE/ASO2184de1A 1 ASOK710X 3 (025) IC XmnI2372de18 1 B PAGE/sequenceExon 15: S945L 1 C TaqlExon 17b:L1065P I MnlIL1077P 1 A ASOY1092X 3 (025) 2C,IA Rsal/ASOExon 19: RI1162X 6 (0-5) 5C,IA DdeI/ASO3659delC 3 (025) 3C ASOExon 20: G1244E 2 2A MboIIS1251N 2 2C RsaI3905insT 4 (0-33) 4C PAGE/ASOW1282X 18 (105) 15B,1D MnlI/ASOR1283K 1 C Mnll/sequenceExon 21: N1303K 22 (1-8) 18B,lA,ID Modified primers+BstNI 47 mutations 1031 (85 9) least one CF chromosome (table): 21 of them are very rare as they were found on only one CF chromosome in our population. Login to comment

PMID: 7521710 [PubMed] Ravnik-Glavac M et al: "Sensitivity of single-strand conformation polymorphism and heteroduplex method for mutation detection in the cystic fibrosis gene."

No. Sentence Comment

121 1078delT (35), L327R (Ravnik-Glavac a al., unpublished), R334W (36), D36K (31), R347L (26), R347P (14), A349V (26), R352Q (30), 1221delCT (34); Exon 8: W401X (31), 1342-1G-C (25); Exon 9: G458V (37), 1525 -1G-A (38); Exon 10: S492F (34), Q493X (39), 1609delCA (40,17), deltaI507 (39,41), deltaF5O8 (3), 1717-1G-A (39,42); Exon 11: G542X (39), S549N, G551D, R553X (43), R553Q (44), A559T (43), R560K (Fine et al., pers. comm.), R560T (39); Exon 12: Y563N (39), 1833delT (Schwartz et al., pers. comm.), P574H (39), 1898 + 1G-C (31), 1898+3A-G (Ferrari et al., pers. comm.); Exon 13: G628R(G-C) (31), Q685X (Firec et al., pers. comm.), K716X (26), L719X (Dork etal., pers. comm.), 2522insC (15), 2556insAT (45), E827X (34); Exon 14a: E831X (Ffrec et al., pers. comm.), R851X (29), 2721delll (31), C866Y (Audrezet et al., pers. comm.); Exon 14b: 2789+5G-A (Highsmith et al., pers. comm.); Exon 15: 2907denT (21), 2991del32 (Dark and TQmmler, pers. comm.), G970R (31); Exon 16: S977P, 3100insA (D6rk et al., pers. comm.); Exon 17a: I1005R (Dork and TQmmler, pers. comm.), 3272-1G-A (46); Exon 17b: H1054D (F6rec et al., pers. comm.), G1061R (Fdrec et al., pers. comm.), 332Oins5, R1066H, A1067T (34), R1066L (Fe"rec etal., pers. comm.), R1070Q (46), E1104X (Zielenski el al., pers. comm.), 3359delCT (46), L1077P (Bozon « a/., pers. comm.), H1085R (46), Y1092X (Bozon etal., pers. comm.), W1098R, M1101K (Zielenski et al., pers. comm.); Exon 18: D1152H (Highsmith et al., pers. comm.); Exon 19:R1162X (36), 3659delC (39), 3662delA (25), 3667del4 (Chillon et al., pers. comm.), 3737ddA (35), 3821ddT (15), I1234V (35), S1235R (31), Q1238X (26), 3849G-A (25), 385O-3T-G (38); Exon20:3860ins31 (Chillon etal., pers. comm.), S1255X (47), 3898insC (26), 3905insT (Malik et al., pers. comm.), D127ON (48), W1282X (49), Q1291R (Dork et al., pers. comm.), Exon 21: N1303H (35), N13O3K (50), W1316X (43); Exon 22: 11328L/4116delA (Dork and TQmmler, pers. comm.), E1371X (25); Exon 23: 4374+ 1G-T (38); Exon 24: 4382delA (Claustres et al., pers. comm.). Login to comment

PMID: 7513293 [PubMed] Chillon M et al: "Analysis of the CFTR gene confirms the high genetic heterogeneity of the Spanish population: 43 mutations account for only 78% of CF chromosomes."

No. Sentence Comment

31 At present, we have not detected any Spanish CF chromosomes bearing any of the following mutations: 394delTA, Y122X, 556delA, 852de122, R347P, $492F, 1677delTA, V520F, Q552X, R553X, L559S, R560K, R560T, Y563N, P564H, 2043delG, 3320ins5, R1066H, A1067T, H1085R, 3732delA, 3737delA, I1234V, S1255P, 3898insC, Q1291H or 4005+ 1G---~A. Login to comment

PMID: 23835419 [PubMed] Loo TW et al: "Corrector VX-809 stabilizes the first transmembrane domain of CFTR."

No. Sentence Comment

64 Arginine point mutations on the predicted aqueous face of TM6 (positions 338, 341, 344, 345, 348, 351) were introduced into processing mutants DF508, G232D, and H1085R. Login to comment
175 By contrast, other CFTR mutants defective in processing such as V232D and H1085R have half-lives similar to wild-type CFTR after rescue [14]. Login to comment
176 To test if other CFTR processing mutants could be rescued by TM6 mutations, arginine mutations were introduced at positions Ile338, Ser341, Ile344, Val345, Met348, and Thr351 of the V232D and H1085R processing mutants. Login to comment
177 None of the introduced arginines were found to promote maturation of V232D or H1085R. Login to comment
178 Examples of typical results obtained with the I344R or M348R mutations introduced into V232D or H1085R are shown in Fig. 5B. Login to comment
179 It was possible that the processing mutations in the TMDs (V232D (TMD1) or H1085R (TMD2)) cannot be rescued by a direct rescue approach using suppressor mutations. Login to comment
180 To test if the V232D or H1085R mutants could be rescued by suppressor mutations in other domains, suppressor mutations in NBD1 (I539T), the NBD1-TMD2 interface (V510D), or TMD2 (R1070W) (only V232D) locations were introduced into the mutants. Login to comment
183 This result shows that the V510D and H1085R mutants could indeed be directly rescued by a suppressor mutation. Login to comment
196 RDR1 differs from VX-809 however, as we find that it does not rescue processing mutations in other domains such as V232D (TMD1) or H1085R (TMD2) (unpublished observations). Login to comment
220 None of the arginines introduced into CFTR processing mutants DF508, V232D, or H1085R promoted maturation. Login to comment
236 (B) Extracts of cells expressing wild-type CFTR or mutants V232D or H1085R with or without the I344R or M348R mutations were subjected to immunoblot analysis. Login to comment
237 (C) Extracts of cells expressing processing mutants DF508, V232D, or H1085R with or without the V510D, I539T, or R1070W suppressor mutations were subjected to immunoblot analysis. Login to comment

PMID: 23891399 [PubMed] Van Goor F et al: "Effect of ivacaftor on CFTR forms with missense mutations associated with defects in protein processing or function."

No. Sentence Comment

42 For example, severe processing mutations such as F508del-, R1066H-, and H1085R-CFTR exhibited a low level of mature CFTR (b5% normal). Login to comment
44 None M1V A46D E56K P67L R74W G85E E92K D110E D110H R117C R117H E193K L206W R334W I336K T338I S341P R347H R347P R352Q A455E L467P S492F F508del V520F A559T R560S R560T A561E Y569D D579G R668C L927P S945L S977F L997F F1052V H1054D K1060T L1065P R1066C R1066H R1066M A1067T R1070Q R1070W F1074L L1077P H1085R M1101K D1152H S1235R D1270N N1303K 0 100 200 300 400 500 600 * * * CFTR Mutation mRNA (% Normal CFTR) Fig. 1. Login to comment
64 Mutant CFTR form CFTR processing Mature/total % Normal CFTR Normal 0.89 &#b1; 0.01 100.0 &#b1; 18.5 G85E -0.05 &#b1; 0.04 -1.0 &#b1; 0.9 R560S 0.00 &#b1; 0.00 0.0 &#b1; 0.0 R1066C 0.02 &#b1; 0.01 0.0 &#b1; 0.0 S492F 0.00 &#b1; 0.00 0.1 &#b1; 0.1 R560T 0.01 &#b1; 0.01 0.2 &#b1; 0.1 V520F 0.05 &#b1; 0.03 0.3 &#b1; 0.2 M1101K 0.05 &#b1; 0.03 0.3 &#b1; 0.1 A561E 0.08 &#b1; 0.04 0.5 &#b1; 0.2 R1066M 0.02 &#b1; 0.02 0.5 &#b1; 0.4 N1303K 0.02 &#b1; 0.02 0.5 &#b1; 0.3 A559T 0.16 &#b1; 0.09 0.6 &#b1; 0.2 M1V 0.06 &#b1; 0.06 0.7 &#b1; 0.6 Y569D 0.11 &#b1; 0.04 0.6 &#b1; 0.2 R1066H 0.08 &#b1; 0.02a 0.7 &#b1; 0.2a L1065P 0.05 &#b1; 0.05 1.0 &#b1; 0.8 L467P 0.10 &#b1; 0.07 1.2 &#b1; 0.8 L1077P 0.08 &#b1; 0.04 1.5 &#b1; 0.6 A46D 0.21 &#b1; 0.08 1.9 &#b1; 0.5a E92K 0.06 &#b1; 0.05 1.9 &#b1; 1.3 H1054D 0.09 &#b1; 0.04 1.9 &#b1; 0.8 F508del 0.09 &#b1; 0.02a 2.3 &#b1; 0.5a H1085R 0.06 &#b1; 0.01a 3.0 &#b1; 0.7a I336K 0.42 &#b1; 0.05a 6.5 &#b1; 0.7a L206W 0.35 &#b1; 0.10a 6.8 &#b1; 1.7a F1074L 0.52 &#b1; 0.03a 10.9 &#b1; 0.6a A455E 0.26 &#b1; 0.10a 11.5 &#b1; 2.5a E56K 0.29 &#b1; 0.04a 12.2 &#b1; 1.5a R347P 0.48 &#b1; 0.04a 14.6 &#b1; 1.8a R1070W 0.61 &#b1; 0.04a 16.3 &#b1; 0.6a P67L 0.36 &#b1; 0.04a 28.4 &#b1; 6.8a R1070Q 0.90 &#b1; 0.01a 29.5 &#b1; 1.4a S977F 0.97 &#b1; 0.01a 37.3 &#b1; 2.4a A1067T 0.78 &#b1; 0.03a 38.6 &#b1; 6.1a D579G 0.72 &#b1; 0.02a 39.3 &#b1; 3.1a D1270N 1.00 &#b1; 0.00a,c 40.7 &#b1; 1.2a S945L 0.65 &#b1; 0.04a 42.4 &#b1; 8.9a L927P 0.89 &#b1; 0.01a,b 43.5 &#b1; 2.5a,b R117C 0.87 &#b1; 0.02a,b 49.1 &#b1; 2.9a,b T338I 0.93 &#b1; 0.03a,b 54.2 &#b1; 3.7a,b L997F 0.90 &#b1; 0.04a,b 59.8 &#b1; 10.4a,b D110H 0.97 &#b1; 0.01a,b 60.6 &#b1; 1.5a,b S341P 0.79 &#b1; 0.02a 65.0 &#b1; 4.9a,b R668C 0.94 &#b1; 0.03a,b 68.5 &#b1; 1.9a,b R74W 0.78 &#b1; 0.01a 69.0 &#b1; 2.7a,b D110E 0.92 &#b1; 0.05a,b 87.5 &#b1; 9.5a,b R334W 0.91 &#b1; 0.05a,b 97.6 &#b1; 10.0a,b K1060T 0.87 &#b1; 0.02a,b 109.9 &#b1; 28.0a,b R347H 0.96 &#b1; 0.02a,c 120.7 &#b1; 2.8a,b S1235R 0.96 &#b1; 0.00a,c 139.0 &#b1; 9.0a,b E193K 0.84 &#b1; 0.02a,b 143.0 &#b1; 17.1a,b R117H 0.86 &#b1; 0.01a,b 164.5 &#b1; 34.2a,b R352Q 0.98 &#b1; 0.01a,b 179.9 &#b1; 8.0a,c F1052V 0.90 &#b1; 0.01a,b 189.9 &#b1; 33.1a,b D1152H 0.96 &#b1; 0.02a,c 312.0 &#b1; 45.5a,b Notes to Table 1: Quantification of steady-state CFTR maturation expressed as the mean (&#b1;SEM; n = 5-9) ratio of mature CFTR to total CFTR (immature plus mature) or level of mature mutant CFTR relative to mature normal-CFTR (% normal CFTR) in FRT cells individually expressing CFTR mutations. Login to comment
74 Because the level of CFTR mRNA was similar across the panel of cell lines tested, the range in baseline activity and ivacaftor response likely reflects the severity of the functional defect and/or the 0 50 100 150 200 S341P R347P L467P S492F A559T A561E Y569D L1065P R1066C R1066M L1077P M1101K N1303K R560S L927P R560T H1085R V520F E92K M1V F508del H1054D I336K A46D G85E R334W T338I R1066H R352Q R117C L206W R347H S977F S945L A455E F1074L E56K P67L R1070W D110H D579G D110E R1070Q L997F A1067T E193K R117H R74W K1060T R668C D1270N D1152H S1235R F1052V Baseline With ivacaftor * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Chloride transport (% Normal) Mutant CFTR form 0 100 200 300 400 S341P R347P L467P S492F A559T A561E Y569D L1065P R1066C R1066M L1077P M1101K N1303K R560S L927P R560T H1085R V520F E92K M1V F508del H1054D I336K A46D G85E R334W T338I R1066H R352Q R117C L206W R347H S977F S945L A455E F1074L P67L E56K R1070W D110H D579G D110E R1070Q L997F A1067T E193K R117H R74W K1060T R668C D1270N D1152H S1235R F1052V * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Mature CFTR (% Normal) Mutant CFTR form A B Fig. 2. Login to comment
82 Mutation Patientsa Chloride transport (bc;A/cm2 ) Chloride transport (% normal) EC50 Baseline With ivacaftor Baseline With ivacaftor Fold increase over baselineb Normal 204.5 &#b1; 33.3 301.3 &#b1; 33.8c 100.0 &#b1; 16.3 147.3 &#b1; 16.5c 1.5 266 &#b1; 42 G551D 1282 1.5 &#b1; 0.7 113.2 &#b1; 13.0c 1.0 &#b1; 0.5 55.3 &#b1; 6.3c 55.3 312 &#b1; 73 F1052V 12 177.3 &#b1; 13.7 410.2 &#b1; 11.3c 86.7 &#b1; 6.7 200.7 &#b1; 5.6c 2.3 177 &#b1; 14 S1235R ND 160.6 &#b1; 25.7 352.1 &#b1; 43.4c 78.5 &#b1; 12.6 172.2 &#b1; 21.2c 2.2 282 &#b1; 104 D1152H 185 117.3 &#b1; 23.0 282.7 &#b1; 46.9c 57.4 &#b1; 11.2 138.2 &#b1; 22.9c 2.4 178 &#b1; 67 D1270N 32 109.5 &#b1; 20.5 209.5 &#b1; 27.4c 53.6 &#b1; 10.0 102.4 &#b1; 13.4c 1.9 254 &#b1; 56 R668C 45 99.0 &#b1; 9.4 217.6 &#b1; 11.7c 48.4 &#b1; 4.6 106.4 &#b1; 5.7c 2.2 517 &#b1; 105 K1060T ND 89.0 &#b1; 9.8 236.4 &#b1; 20.3c 43.5 &#b1; 4.8 115.6 &#b1; 9.9c 2.7 131 &#b1; 73 R74W 25 86.8 &#b1; 26.9 199.1 &#b1; 16.8c 42.5 &#b1; 13.2 97.3 &#b1; 8.2c 2.3 162 &#b1; 17 R117H 739 67.2 &#b1; 13.3 274.1 &#b1; 32.2c 32.9 &#b1; 6.5 134.0 &#b1; 15.7c 4.1 151 &#b1; 14 E193K ND 62.2 &#b1; 9.8 379.1 &#b1; 1.1c 30.4 &#b1; 4.8 185.4 &#b1; 1.0c 6.1 240 &#b1; 20 A1067T ND 55.9 &#b1; 3.2 164.0 &#b1; 9.7c 27.3 &#b1; 1.6 80.2 &#b1; 4.7c 2.9 317 &#b1; 214 L997F 27 43.7 &#b1; 3.2 145.5 &#b1; 4.0c 21.4 &#b1; 1.6 71.2 &#b1; 2.0c 3.3 162 &#b1; 12 R1070Q 15 42.0 &#b1; 0.8 67.3 &#b1; 2.9c 20.6 &#b1; 0.4 32.9 &#b1; 1.4c 1.6 164 &#b1; 20 D110E ND 23.3 &#b1; 4.7 96.4 &#b1; 15.6c 11.4 &#b1; 2.3 47.1 &#b1; 7.6c 4.1 213 &#b1; 51 D579G 21 21.5 &#b1; 4.1 192.0 &#b1; 18.5c 10.5 &#b1; 2.0 93.9 &#b1; 9.0c 8.9 239 &#b1; 48 D110H 30 18.5 &#b1; 2.2 116.7 &#b1; 11.3c 9.1 &#b1; 1.1 57.1 &#b1; 5.5c 6.2 249 &#b1; 59 R1070W 13 16.6 &#b1; 2.6 102.1 &#b1; 3.1c 8.1 &#b1; 1.3 49.9 &#b1; 1.5c 6.2 158 &#b1; 48 P67L 53 16.0 &#b1; 6.7 88.7 &#b1; 15.7c 7.8 &#b1; 3.3 43.4 &#b1; 7.7c 5.6 195 &#b1; 40 E56K ND 15.8 &#b1; 3.1 63.6 &#b1; 4.4c 7.7 &#b1; 1.5 31.1 &#b1; 2.2c 4.0 123 &#b1; 33 F1074L ND 14.0 &#b1; 3.4 43.5 &#b1; 5.4c 6.9 &#b1; 1.6 21.3 &#b1; 2.6c 3.1 141 &#b1; 19 A455E 120 12.9 &#b1; 2.6 36.4 &#b1; 2.5c 6.3 &#b1; 1.2 17.8 &#b1; 1.2c 2.8 170 &#b1; 44 S945L 63 12.3 &#b1; 3.9 154.9 &#b1; 47.6c 6.0 &#b1; 1.9 75.8 &#b1; 23.3c 12.6 181 &#b1; 36 S977F 9 11.3 &#b1; 6.2 42.5 &#b1; 19.1c 5.5 &#b1; 3.0 20.8 &#b1; 9.3c 3.8 283 &#b1; 36 R347H 65 10.9 &#b1; 3.3 106.3 &#b1; 7.6c 5.3 &#b1; 1.6 52.0 &#b1; 3.7c 9.8 280 &#b1; 35 L206W 81 10.3 &#b1; 1.7 36.4 &#b1; 2.8c 5.0 &#b1; 0.8 17.8 &#b1; 1.4c 3.6 101 &#b1; 13 R117C 61 5.8 &#b1; 1.5 33.7 &#b1; 7.8c 2.9 &#b1; 0.7 16.5 &#b1; 3.8c 5.7 380 &#b1; 136 R352Q 46 5.5 &#b1; 1.0 84.5 &#b1; 7.8c 2.7 &#b1; 0.5 41.3 &#b1; 3.8c 15.2 287 &#b1; 75 R1066H 29 3.0 &#b1; 0.3 8.0 &#b1; 0.8c 1.5 &#b1; 0.1 3.9 &#b1; 0.4c 2.6 390 &#b1; 179 T338I 54 2.9 &#b1; 0.8 16.1 &#b1; 2.4c 1.4 &#b1; 0.4 7.9 &#b1; 1.2c 5.6 334 &#b1; 38 R334W 150 2.6 &#b1; 0.5 10.0 &#b1; 1.4c 1.3 &#b1; 0.2 4.9 &#b1; 0.7c 3.8 259 &#b1; 103 G85E 262 1.6 &#b1; 1.0 1.5 &#b1; 1.2 0.8 &#b1; 0.5 0.7 &#b1; 0.6 NS NS A46D ND 2.0 &#b1; 0.6 1.1 &#b1; 1.1 1.0 &#b1; 0.3 0.5 &#b1; 0.6 NS NS I336K 29 1.8 &#b1; 0.2 7.4 &#b1; 0.1c 0.9 &#b1; 0.1 3.6 &#b1; 0.1c 4 735 &#b1; 204 H1054D ND 1.7 &#b1; 0.3 8.7 &#b1; 0.3c 0.8 &#b1; 0.1 4.2 &#b1; 0.1c 5.3 187 &#b1; 20 F508del 29,018 0.8 &#b1; 0.6 12.1 &#b1; 1.7c 0.4 &#b1; 0.3 5.9 &#b1; 0.8c 14.8 129 &#b1; 38 M1V 9 0.7 &#b1; 1.4 6.5 &#b1; 1.9c 0.4 &#b1; 0.7 3.2 &#b1; 0.9c 8.0 183 &#b1; 85 E92K 14 0.6 &#b1; 0.2 4.3 &#b1; 0.8c 0.3 &#b1; 0.1 2.1 &#b1; 0.4c 7.0 198 &#b1; 46 V520F 58 0.4 &#b1; 0.2 0.5 &#b1; 0.2 0.2 &#b1; 0.1 0.2 &#b1; 0.1 NS NS H1085R ND 0.3 &#b1; 0.2 2.1 &#b1; 0.4 0.2 &#b1; 0.1 1.0 &#b1; 0.2 NS NS R560T 180 0.3 &#b1; 0.3 0.5 &#b1; 0.5 0.1 &#b1; 0.1 0.2 &#b1; 0.2 NS NS L927P 15 0.2 &#b1; 0.1 10.7 &#b1; 1.7c 0.1 &#b1; 0.1 5.2 &#b1; 0.8c 52.0 313 &#b1; 66 R560S ND 0.0 &#b1; 0.1 -0.2 &#b1; 0.2 0.0 &#b1; 0.0 -0.1 &#b1; 0.1 NS NS N1303K 1161 0.0 &#b1; 0.0 1.7 &#b1; 0.3 0.0 &#b1; 0.0 0.8 &#b1; 0.2 NS NS M1101K 79 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS L1077P 42 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS R1066M ND 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS R1066C 100 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS L1065P 25 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS Y569D 9 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS A561E ND 0.0 &#b1; 0.1 0.0 &#b1; 0.1 0.0 &#b1; 0.0 0.0 &#b1; 0.1 NS NS A559T 43 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS S492F 16 0.0 &#b1; 0.0 1.7 &#b1; 1.2 0.0 &#b1; 0.0 0.8 &#b1; 0.6 NS NS L467P 16 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS R347P 214 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS S341P 9 0.0 &#b1; 0.0 0.2 &#b1; 0.2 0.0 &#b1; 0.0 0.1 &#b1; 0.1 NS NS a Number of individuals with the individual mutation in the CFTR-2 database (www.CFTR2.org). Login to comment

PMID: 24412276 [PubMed] Loo TW et al: "The cystic fibrosis V232D mutation inhibits CFTR maturation by disrupting a hydrophobic pocket rather than formation of aberrant interhelical hydrogen bonds."

No. Sentence Comment

169 For example, V510D promotes maturation of mutants with processing mutations in TMD1 (V232D), TMD2 (H1085R) and NBD1 (DF508) whereas other suppressors such as I539T and R1070W promote maturation of DF508 CFTR but not mutants V232D or H1085R [19]. Login to comment
258 Cross-linking analysis suggests that V232D causes incomplete packing of the TM segments Previous studies on processing mutations in NBD1 (DF508) or in the fourth intracellular loop connecting TM segments 10 and 11 (Q1071P or H1085R) showed that they trapped CFTR at an early folding step resulting in incomplete packing of the TM segments [17,50]. Login to comment
313 We predict that V232D inhibits CFTR maturation by a mechanism that is similar to that proposed for the Q1071P, H1085R [50] and DF508 [17,26] processing mutations (see Fig. 11). Login to comment