ABCMdb

ABCC1 p.His1486Asn

Predicted by SNAP2:	A: D (85%), C: D (85%), D: D (91%), E: D (91%), F: D (91%), G: D (85%), I: D (91%), K: D (85%), L: D (91%), M: D (85%), N: D (80%), P: D (91%), Q: D (80%), R: D (85%), S: D (80%), T: D (85%), V: D (85%), W: D (91%), Y: D (85%),
Predicted by PROVEAN:	A: D, C: D, D: D, E: D, F: D, G: D, I: D, K: D, L: D, M: D, N: D, P: D, Q: D, R: D, S: D, T: D, V: D, W: D, Y: D,

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Publications
[hide] Hydrogen-bond formation of the residue in H-loop o... Biochim Biophys Acta. 2007 Feb;1768(2):324-35. Epub 2006 Nov 18. Yang R, Chang XB
Hydrogen-bond formation of the residue in H-loop of the nucleotide binding domain 2 with the ATP in this site and/or other residues of multidrug resistance protein MRP1 plays a crucial role during ATP-dependent solute transport.
Biochim Biophys Acta. 2007 Feb;1768(2):324-35. Epub 2006 Nov 18., [PMID:17187755]

Abstract [show]
MRP1 couples ATP binding/hydrolysis to solute transport. We have shown that ATP binding to nucleotide-binding-domain 1 (NBD1) plays a regulatory role whereas ATP hydrolysis at NBD2 plays a crucial role in ATP-dependent solute transport. However, how ATP is hydrolyzed at NBD2 is not well elucidated. To partially address this question, we have mutated the histidine residue in H-loop of MRP1 to either a residue that prevents the formation of hydrogen-bonds with ATP and other residues in MRP1 or a residue that may potentially form these hydrogen-bonds. Interestingly, substitution of H827 in NBD1 with residues that prevented formation of these hydrogen-bonds had no effect on the ATP-dependent solute transport whereas corresponding mutations in NBD2 almost abolished the ATP-dependent solute transport completely. In contrast, substitutions of H1486 in H-loop of NBD2 with residues that might potentially form these hydrogen-bonds exerted either full function or partial function, implying that hydrogen-bond formation between the residue at 1486 and the gamma-phosphate of the bound ATP and/or other residues, such as putative catalytic base E1455, together with S769, G771, T1329 and K1333, etc., holds all the components necessary for ATP binding/hydrolysis firmly so that the activated water molecule can efficiently hydrolyze the bound ATP at NBD2.

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No. Sentence Comment
43 In addition, the following mutations as shown in Fig. 5A (H827D, H1486D, H827D/ H1486D, H827N, H1486N, H827N/H1486N, H827E, H1486E, H827E/ H1486E, H827Q, H1486Q, H827Q/H1486Q, H827Y, H1486Y, H827Y/ H1486Y, H827W, H1486W and H827W/H1486W) were also introduced into the full length MRP1 cDNA by using the following primers: H827D/forward, 5'-CGG ATC TTG GTC ACG GAC AGC ATG AGC TAC TTG-3'; H827D/ reverse, 5'-CAA GTA GCT CAT GCT GTC CGT GAC CAA GAT CCG-3'; H1486D/forward, 5'-GTC CTC ACC ATC GCC GAC CGG CTC AAC ACC ATC-3'; H1486D/reverse, 5'-GAT GGT GTT GAG CCG GTC GGC GAT GGT GAG GAC-3'; H827N/forward, 5'-CGG ATC TTG GTC ACG AAC AGC ATG AGC TAC TTG-3'; H827N/reverse, 5'-CAA GTA GCT CAT GCT GTT CGT GAC CAA GAT CCG-3'; H1486N/forward, 5'-GTC CTC ACC ATC GCC AAC CGG CTC AAC ACC ATC-3'; H1486N/reverse, 5'-GAT GGT GTT GAG CCG GTT GGC GAT GGT GAG GAC-3'; H827E/forward, 5'-CGG ATC TTG GTC ACG GAG AGC ATG AGC TAC TTG-3'; H827E/reverse, 5'-CAA GTA GCT CAT GCT CTC CGT GAC CAA GAT CCG-3'; H1486E/forward, 5'-GTC CTC ACC ATC GCC GAG CGG CTC AAC ACC ATC-3'; H1486E/reverse, 5'-GAT GGT GTT GAG CCG CTC GGC GAT GGT GAG GAC-3'; H827Q/forward, 5'- CGG ATC TTG GTC ACG CAG AGC ATG AGC TAC TTG-3'; H827Q/ reverse, 5'-CAA GTA GCT CAT GCT CTG CGT GAC CAA GAT CCG-3'; H1486Q/forward, 5'-GTC CTC ACC ATC GCC CAG CGG CTC AAC ACC ATC-3'; H1486Q/reverse, 5'-GAT GGT GTT GAG CCG CTG GGC GAT GGT GAG GAC-3'; H827Y/forward, 5'-CGG ATC TTG GTC ACG TAC AGC ATG AGC TAC TTG-3'; H827Y/reverse, 5'-CAA GTA GCT CAT GCT GTA CGT GAC CAA GAT CCG-3'; H1486Y/forward, 5'-GTC CTC ACC ATC GCC TAC CGG CTC AAC ACC ATC-3'; H1486Y/reverse, 5'-GAT GGT GTT GAG CCG GTA GGC GAT GGT GAG GAC-3'; H827W/forward, 5'-CGG ATC TTG GTC ACG TGG AGC ATG AGC TAC TTG-3'; H827W/reverse, 5'-CAA GTA GCT CAT GCT CCA CGT GAC CAA GAT CCG-3'; H1486W/forward, 5'-GTC CTC ACC ATC GCC TGG CGG CTC AAC ACC ATC-3'; H1486W/reverse, 5'-GAT GGT GTT GAG CCG CCA GGC GAT GGT GAG GAC-3'. Login to comment
164 In order to test this hypothesis, H827D, H1486D, H827D/H1486D, H827N, H1486N, H827N/H1486N, H827E, H1486E, H827E/H1486E, H827Q, H1486Q, H827Q/H1486Q, H827Y, H1486Y, H827Y/ H1486Y, H827W, H1486W and H827W/H1486W mutations (Fig. 5A) were introduced into full length of MRP1 cDNA in pNUT/MRP1/His and expressed in BHK cells at 37 °C. Login to comment
169 H1486N had ~60% of wild-type MRP1 transport activity, whereas H1486D dropped this activity to ~20%. Login to comment
210 In contrast, mutations in NBD2, such as H1486L, H1486F, H1486D, H1486N, H1486E, H1486Q, H1486Y and H1486W, have variant effects on the ATP-dependent LTC4 transport (Figs. 1C, 4B and 5C)). Login to comment
224 However, H1486E exerts ~36% of wild-type MRP1 transport activity (Fig. 5C), presumably the carboxyl group of E1486 cannot form strong hydrogen-bonds with the γ-phosphate of the bound ATP and the putative catalytic base E1455 in NBD2. Accordingly, although H1486N has the same side group as the H1486Q mutation, except that the length of the side chain in H1486N is one carbon (or 1.541 Å) shorter than in H1486Q, the transport activity of H1486N dropped from ~112% (H1486Q) to ~60% (Fig. 5C), implying that the distance between two groups forming hydrogen-bond also plays an important role. Login to comment