ABCMdb

ABCB1 p.Asp164Gln

Predicted by SNAP2:	A: D (75%), C: D (75%), E: D (66%), F: D (80%), G: D (80%), H: D (80%), I: D (85%), K: D (85%), L: D (85%), M: D (80%), N: D (75%), P: D (91%), Q: D (75%), R: D (85%), S: D (71%), T: D (75%), V: D (80%), W: D (85%), Y: D (75%),
Predicted by PROVEAN:	A: D, C: D, E: D, F: D, G: D, H: 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] A mutation within the extended X loop abolished su... J Biol Chem. 2015 Feb 20;290(8):4896-907. doi: 10.1074/jbc.M114.588566. Epub 2014 Dec 22. Kluth M, Stindt J, Droge C, Linnemann D, Kubitz R, Schmitt L
A mutation within the extended X loop abolished substrate-induced ATPase activity of the human liver ATP-binding cassette (ABC) transporter MDR3.
J Biol Chem. 2015 Feb 20;290(8):4896-907. doi: 10.1074/jbc.M114.588566. Epub 2014 Dec 22., [PMID:25533467]

Abstract [show]
The human multidrug resistance protein 3 (MDR3/ABCB4) belongs to the ubiquitous family of ATP-binding cassette (ABC) transporters and is located in the canalicular membrane of hepatocytes. There it flops the phospholipids of the phosphatidylcholine (PC) family from the inner to the outer leaflet. Here, we report the characterization of wild type MDR3 and the Q1174E mutant, which was identified previously in a patient with progressive familial intrahepatic cholestasis type 3 (PFIC-3). We expressed different variants of MDR3 in the yeast Pichia pastoris, purified the proteins via tandem affinity chromatography, and determined MDR3-specific ATPase activity in the presence or absence of phospholipids. The ATPase activity of wild type MDR3 was stimulated 2-fold by liver PC or 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine lipids. Furthermore, the cross-linking of MDR3 with a thiol-reactive fluorophore blocked ATP hydrolysis and exhibited no PC stimulation. Similarly, phosphatidylethanolamine, phosphatidylserine, and sphingomyelin lipids did not induce an increase of wild type MDR3 ATPase activity. The phosphate analogues beryllium fluoride and aluminum fluoride led to complete inhibition of ATPase activity, whereas orthovanadate inhibited exclusively the PC-stimulated ATPase activity of MDR3. The Q1174E mutation is located in the nucleotide-binding domain in direct proximity of the leucine of the ABC signature motif and extended the X loop, which is found in ABC exporters. Our data on the Q1174E mutant demonstrated basal ATPase activity, but PC lipids were incapable of stimulating ATPase activity highlighting the role of the extended X loop in the cross-talk of the nucleotide-binding domain and the transmembrane domain.

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Sentences [show]
No. Sentence Comment
310 They demonstrated that the amino acid residue Gln-1175 of NBD2, which is identical to Gln-1174 in MDR3, hydrogen bonds with Asp-164 (Asp-166 in MDR3) of the ICL1 and identified this residue pair Asp-164-Gln-1175 as key residue pair in the transmission interface. Login to comment
309 They demonstrated that the amino acid residue Gln-1175 of NBD2, which is identical to Gln-1174 in MDR3, hydrogen bonds with Asp-164 (Asp-166 in MDR3) of the ICL1 and identified this residue pair Asp-164-Gln-1175 as key residue pair in the transmission interface. Login to comment