title: Identification of an amino acid residue in ATP-binding cassette transport G1 critical for mediating cholesterol efflux.
abstract: The ATP-binding cassette transporter G1 (ABCG1) mediates free cholesterol efflux onto lipidated apolipoprotein A-I (apoA-I) and plays an important role in macrophage reverse cholesterol transport thereby reducing atherosclerosis. However, how ABCG1 mediates the efflux of cholesterol onto lipidated apoA-I is unclear. Since the crystal structure of ABCG family is not available, other approaches such as site-directed mutagenesis have been widely used to identify amino acid residues important for protein functions. We noticed that ABCG1 contains a single cysteine residue in its putative transmembrane domains. This cysteine residue locates at position 514 (Cys(514)) within the third putative transmembrane domain and is highly conserved. Replacement of Cys(514) with Ala (C514A) essentially abolished ABCG1-mediated cholesterol efflux onto lipidated apoA-I. Substitution of Cys(514) with more conserved amino acid residues, Ser or Thr, also significantly decreased cholesterol efflux. However, mutation C514A had no detectable effect on protein stability and trafficking. Mutation C514A also did not affect the dimerization of ABCG1. Our findings demonstrated that the sulfhydryl group of Cys residue located at position 514 plays a critical role in ABCG1-mediated cholesterol efflux. This article is part of a Special Issue entitled Advances in High Density Lipoprotein Formation and Metabolism: A Tribute to John F. Oram (1945-2010).
authors.affiliation: Department of Pediatrics and Group on the Molecular and Cell Biology of Lipids, University of Alberta, Edmonton, Alberta, Canada T6G 2S2.
authors.forename: Xia
authors.initials: X
authors.lastname: Gao
authors.forename: Hongmei
authors.initials: H
authors.lastname: Gu
authors.forename: Ge
authors.initials: G
authors.lastname: Li
authors.forename: Kerry-Anne
authors.initials: KA
authors.lastname: Rye
authors.forename: Da-Wei
authors.initials: DW
authors.lastname: Zhang
mesh_terms: D000070998:ATP Binding Cassette Transporter, Subfamily G, Member 1; D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D016632:Apolipoprotein A-I / Q000378:metabolism; D037281:Calnexin / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism*; D002850:Chromatography, Gel; D017124:Conserved Sequence; D003545:Cysteine / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D021381:Protein Transport
chemical_list: C419353:ABCG1 protein, human; C505693:APOA1 protein, human; D000070998:ATP Binding Cassette Transporter, Subfamily G, Member 1; D018528:ATP-Binding Cassette Transporters; D016632:Apolipoprotein A-I; D037281:Calnexin; D002784:Cholesterol; D003545:Cysteine