ABCMdb

ABCC2 p.Pro1158Ala

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Publications

PMID: 17494643 [PubMed] Letourneau IJ et al: "Mutational analysis of a highly conserved proline residue in MRP1, MRP2, and MRP3 reveals a partially conserved function."

No. Sentence Comment

5 As observed for MRP1-Pro1150Ala, LTC4 transport by MRP2-Pro1158Ala was decreased. Login to comment
122 As shown in Fig. 2, all three Pro to Ala mutants (MRP1-Pro1150Ala, MRP2-Pro1158Ala, and MRP3-Pro1147Ala) were expressed at levels comparable to those of their corresponding wild-type proteins, confirming that the Pro residue at this position is not required for expression of the MRP proteins in the plasma membrane of mammalian cells. Login to comment
141 Protein expression levels of mutants MRP1-Pro1150Ala, MRP2-Pro1158Ala, and MRP3-Pro1147Ala and their corresponding wild-type proteins. Login to comment
148 Vesicular transport of 3 H-labeled organic anions by mutants MRP1-Pro1150Ala, MRP2-Pro1158Ala, and MRP3-Pro1147Ala. Login to comment
165 Mutations MRP1-Pro1150Ala, MRP2-Pro1158Ala, and MRP3-Pro1147Ala Do Not Affect [␥-32 P]8N3ATP Binding but Decrease Vanadate-Induced Trapping of [␣-32 P]8N3ADP. Login to comment
175 Thus, 8N3ADP trapping by the MRP3 mutant was decreased as observed with the MRP1-Pro1150Ala and MRP2-Pro1158Ala mutants, although under different conditions. Login to comment
208 Thus, although 8N3ATP binding to MRP1-Pro1150Ala, MRP2-Pro1158Ala, and MRP3-Pro1147Ala remained unchanged, the level of vanadate-induced 8N3ADP trapping was very low for all three mutant transporters. Login to comment
59 The template for generating the MRP2-Pro1158Ala mutant was created by subcloning a 2.2-kb ApaI/ClaI fragment into pGEM-7Zf(ϩ) (Promega, Madison, WI). Login to comment
62 Mutagenesis was performed according to the manufacturer`s instructions with the following sense primers (substituted nucleotides are underlined, and in both cases, the substitution created a BstUI restriction site, which was used to confirm the successful introduction of the nucleotide substitutions): MRP2-Pro1158Ala, 5Ј-C ACC AGG TCC GCG ATC TAC TCT C-3Ј (BstUI) and MRP3-Pro1147Ala, 5Ј-GTC AGC CGC TCC GCG ATC TAC TCC C-3Ј (BstUI). Login to comment
63 The MRP2-Pro1158Ala mutation was subcloned back into pcDNA3.1(-) MRP2 as a 1.75-kb Bsu36I/SfiI fragment and the MRP3-Pro1147Ala mutation was subcloned into pcDNA3.1(ϩ) MRP3 as a 1.24-kb AgeI/SacII fragment (Oleschuk et al., 2003). Login to comment
120 To determine whether this expression was also true for the corresponding MRP2-Pro1158Ala and MRP3-Pro1147Ala mutants, these mutations were created in pcDNA3.1(Ϯ)-based MRP2 and MRP3 expression vectors and then transfected into HEK293T cells. Login to comment
130 For MRP2-Pro1158Ala, LTC4 transport was also decreased by ϳ50% whereas it was significantly increased (1.8-fold) for MRP3-Pro1147Ala (Fig. 3A). Login to comment
137 In contrast to LTC4 uptake, E217betaG transport by MRP2 and MRP3 was unaffected by the Pro1158Ala and Pro1147Ala mutations, respectively (Fig. 3B). Login to comment
140 In contrast, MTX transport by the MRP2-Pro1158Ala mutant remained the same as that of wild-type MRP2 (Fig. 3C), whereas FIG. 2. Login to comment
160 To determine whether the reduced LTC4 transport by MRP2-Pro1158Ala was associated with reduced substrate binding, [3 H]LTC4 photolabeling of this mutant was compared with that of wild-type MRP2. Login to comment
163 Similarly, MRP2-Pro1158Ala was photolabeled by [3 H]LTC4 to the same extent as wild-type MRP2 despite the reduced LTC4 transport activity of the mutant (Fig. 4B). Login to comment
167 When MRP2-Pro1158Ala and MRP3-Pro1147Ala were photolabeled with [␥-32 P]8N3ATP, the intensity of the signals observed with the mutant and wild-type proteins was also comparable (Fig. 5), suggesting that ATP can bind to the Pro mutants as well as it does to their wild-type counterparts. Login to comment
170 When the MRP2-Pro1158Ala and MRP3-Pro1147Ala mutants were examined under the same conditions, a significant decrease (50%) in the level of vanadate-induced 8N3ADP trapping by MRP2-Pro1158Ala was also observed (Fig. 6B), whereas no trapping of nucleotide was detected with either wild-type MRP3 or its Pro1147Ala mutant (data not shown). Login to comment

PMID: 20082599 [PubMed] Jemnitz K et al: "ABCC2/Abcc2: a multispecific transporter with dominant excretory functions."

No. Sentence Comment

97 Mutant Predicted location Substrate Activity changes Reference Human MRP2 Δ1-188 TMD0 LTC4 ↓ Fernandez et al., 2002 K316A JC, TM6 GMF ↔ Ryu et al., 2000 K324A TM6 GMF ↓ Ryu et al., 2000 K329A TM6 GMF ↔ Ryu et al., 2000 R412G DJ IC MTX ↓ Hulot et al., 2005 W417I IC, TM7-TM8 E2-17βG ↓ Hirouchi et al., 2004 LTC4 ↓ DNP-SG ↓ H439A TM8 GMF ↔ Ryu et al., 2000 K483A IC, JM, TM9 GMF ↓ Ryu et al., 2000 K590A JC, TM11 GMF ↔ Ryu et al., 2000 S789F NBD1 E2-17βG ↓ Hirouchi et al., 2004 LTC4 ↓ DNP-SG ↓↓ R1023A EC, JM, TM13 GMF ↔ Ryu et al., 2000 H1042A TM13 GMF ↔ Ryu et al., 2000 R1100A JC, TM14 GMF ↔ Ryu et al., 2000 P1158A IC, JM, TM15 LTC4 ↓↓ Letourneau et al., 2007 E2-17βG ↔ MTX ↔ Table 1. continued on next page Mutant Predicted location Substrate Activity changes Reference I1173F DJ IC, TM15-16 LTC4 No act Keitel et al., 2003 E2-17βG No act R1210A EC, JC, TM16 GMF ↓↓ Ryu et al., 2000 R1230A TM16 GMF ↔ R1257A JC, TM17 GMF ↓↓ W1254A JC, TM17 E2-17βG ↓↓ Ito et al., 2001a W1254C ↓↓ W1254F ↔ W1254Y ↔ W1254A JC, TM17 LTC4 ↓↓ Ito et al., 2001b W1254C ↓↓↓ W1254F ↓↓ W1254Y ↓↓ W1254A JC, TM17 MTX ↓↓ Ito et al., 2001a W1254C ↓↓ W1254F ↓↓ W1254Y ↓↓↓ A1450T NBD2 E2-17βG ↓↓ Hirouchi et al., 2004 LTC4 ↓↓ DNP-SG ↓↓ Rat Mrp2 K308M IC, JM, TM6 TLC-S ↔ Ito et al., 2001b DNP-G ↑ LTC4 ↓ E3040G ↔ K320M TM6 TLC-S ↑ DNP-G ↑ LTC4 ↓ E3040G ↑ K325M TM6 TLC-S ↓* DNP-G ↓↓↓* LTC4 ↓↓↓* E3040G ↓ D329N TM6 TLC-S ↔ DNP-G ↓ LTC4 ↓↓↓* E3040G ↓ R586L TM11 TLC-S ↓ DNP-G ↓↓* LTC4 ↓↓* E3040G ↔ R1019M IC, JM, TM13 TLC-S ↔ DNP-G ↑* LTC4 ↔ E3040G ↔ R1096L TM14 TLC-S ↑ DNP-G ↑ LTC4 ↔ E3040G ↔ EC, extracellular; IC, intracellular; JC, near the cytosol in the membrane; JM, juxtamembrane; TLC-S, tauro-litocholate-sulfate; GMF, glutathione- methyl-fluorescein; ↑, activity over control>1.2; ↔, 1.2>activity over control>0.8; ↓, 0.8>activity over control>0.5; ↓↓, 0.5>activity over control>0.1; ↓↓↓, 0.1>activity over control. Login to comment