ABCMdb

ABCA1 p.Cys75Ser

Predicted by SNAP2:	A: D (75%), D: D (95%), E: D (95%), F: D (91%), G: D (91%), H: D (85%), I: D (91%), K: D (95%), L: D (91%), M: D (91%), N: D (91%), P: D (95%), Q: D (85%), R: D (95%), S: D (85%), T: D (85%), V: D (91%), W: D (95%), Y: D (91%),
Predicted by PROVEAN:	A: D, D: 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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[hide] Formation of two intramolecular disulfide bonds is... J Biol Chem. 2009 Apr 24;284(17):11293-300. Epub 2009 Mar 3. Hozoji M, Kimura Y, Kioka N, Ueda K
Formation of two intramolecular disulfide bonds is necessary for ApoA-I-dependent cholesterol efflux mediated by ABCA1.
J Biol Chem. 2009 Apr 24;284(17):11293-300. Epub 2009 Mar 3., [PMID:19258317]

Abstract [show]
ABCA1 plays a major role in cholesterol homeostasis and high density lipoprotein (HDL) metabolism. ABCA1 contains disulfide bond(s) between its N- and C-terminal halves, but it remains unclear whether disulfide bond formation is important for the functions of ABCA1 and which cysteines are involved in disulfide bond formation. To answer these questions, we constructed >30 ABCA1 mutants in which 16 extracellular domain (ECD) cysteines were replaced with serines and examined disulfide bond formation, apoA-I binding, and HDL formation in these mutants. From the single cysteine replacements, two cysteines (Cys(75) and Cys(309)) in ECD1 were found to be essential for apoA-I binding. In contrast, in ECD2, only Cys(1477) was found to be essential for HDL formation, and no single cysteine replacement impaired apoA-I binding. The concurrent replacement of two cysteines, Cys(1463) and Cys(1465), impaired apoA-I binding and HDL formation, suggesting that four of five extracellular cysteines (Cys(75), Cys(309), Cys(1463), Cys(1465), and Cys(1477)) are involved in these functions of ABCA1. Trypsin digestion experiments suggested that one disulfide bond is not sufficient and that two intramolecular disulfide bonds (between Cys(75) and Cys(309) in ECD1 and either Cys(1463) or Cys(1465) and Cys(1477) in ECD2) are required for ABCA1 to be fully functional.

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124 TABLE 1 Cysteine mutants of ABCA1 Protein ApoA-I binding Cholesterol efflux Mutations Wild type ϩ ϩ None C1S ϩ ϩ C54S C2S - - C75S C3S ϩ ϩ C81S C4S ϩ ϩ C195S C5S ϩ ϩ C215S C6S - - C309S C7S ϩ ϩ C355S C8S ϩ ϩ C504S C9S ϩ ϩ C626S C10S ϩ ϩ C791S C11S ϩ ϩ C1418S C12S ϩ ϩ C1429S C13S ϩ ϩ C1463S C14S ϩ ϩ C1465S C15S ϩ - C1477S C16S ϩ ϩ C1814S C2/6S - - C75S/C309S C11/12S ϩ ϩ C1418S/C1429S C13/14S - - C1463S/C1465S C13/14/15S - - C1463S/C1465S/C1477S C11/12/13/14S - - C1418S/C1429S/C1463S/C1465S C11/12/13/15S - - C1418S/C1429S/C1463S/C1477S C11/12/14/15S - - C1418S/C1429S/C1465S/C1477S C11/13/14/15S - - C1418S/C1463S/C1465S/C1477S C12/13/14/15S - - C1429S/C1463S/C1465S/C1477S C11/12/13/14/15S - - C1418S/C1429S/C1463S/C1465S/C1477S C2/6/13/14/15S - - C75S/C309S/C1463S/C1465S/C1477S Cys4#1 ϩ ϩ C54S/C81S/C195S/C215S/C355S/C504S/C626S/C791S/C1418S/C1429S/C1465S/1814S Cys4#2 ϩ ϩ C54S/C81S/C195S/C215S/C355S/C504S/C626S/C791S/C1418S/C1429S/C1463S/1814S Cys5 ϩ ϩ C54S/C81S/C195S/C215S/C355S/C504S/C626S/C791S/C1418S/C1429S/C1814S irreversibly abolished apoA-I binding to cells expressing ABCA1(Cys5)-GFP (supplemental Fig. 4). Login to comment
122 TABLE 1 Cysteine mutants of ABCA1 Protein ApoA-I binding Cholesterol efflux Mutations Wild type af9; af9; None C1S af9; af9; C54S C2S afa; afa; C75S C3S af9; af9; C81S C4S af9; af9; C195S C5S af9; af9; C215S C6S afa; afa; C309S C7S af9; af9; C355S C8S af9; af9; C504S C9S af9; af9; C626S C10S af9; af9; C791S C11S af9; af9; C1418S C12S af9; af9; C1429S C13S af9; af9; C1463S C14S af9; af9; C1465S C15S af9; afa; C1477S C16S af9; af9; C1814S C2/6S afa; afa; C75S/C309S C11/12S af9; af9; C1418S/C1429S C13/14S afa; afa; C1463S/C1465S C13/14/15S afa; afa; C1463S/C1465S/C1477S C11/12/13/14S afa; afa; C1418S/C1429S/C1463S/C1465S C11/12/13/15S afa; afa; C1418S/C1429S/C1463S/C1477S C11/12/14/15S afa; afa; C1418S/C1429S/C1465S/C1477S C11/13/14/15S afa; afa; C1418S/C1463S/C1465S/C1477S C12/13/14/15S afa; afa; C1429S/C1463S/C1465S/C1477S C11/12/13/14/15S afa; afa; C1418S/C1429S/C1463S/C1465S/C1477S C2/6/13/14/15S afa; afa; C75S/C309S/C1463S/C1465S/C1477S Cys4#1 af9; af9; C54S/C81S/C195S/C215S/C355S/C504S/C626S/C791S/C1418S/C1429S/C1465S/1814S Cys4#2 af9; af9; C54S/C81S/C195S/C215S/C355S/C504S/C626S/C791S/C1418S/C1429S/C1463S/1814S Cys5 af9; af9; C54S/C81S/C195S/C215S/C355S/C504S/C626S/C791S/C1418S/C1429S/C1814S Two Intramolecular Disulfide Bonds in ABCA1 APRIL 24, 2009ߦVOLUME 284ߦNUMBER 17 JOURNAL OF BIOLOGICAL CHEMISTRY 11297 irreversibly abolished apoA-I binding to cells expressing ABCA1(Cys5)-GFP (supplemental Fig. 4). Login to comment