title: C323 of SR-BI is required for SR-BI-mediated HDL binding and cholesteryl ester uptake.
abstract: Scavenger receptor BI (SR-BI) is an HDL receptor. It binds HDL and mediates the uptake of cholesteryl ester from HDL. Early studies have pointed out that the extracellular domain of SR-BI is critical for SR-BI-mediated cholesteryl ester uptake. However, the extracellular loop of SR-BI is large: it contains 403 amino acids. The HDL binding site and the modulation of SR-BI-mediated cholesteryl ester uptake remain to be identified. In this study, using C323G mutant SR-BI, we showed that C323G mutant SR-BI lost its HDL binding and cholesteryl ester uptake activity, indicating that the highly conserved C323 is required for SR-BI-mediated HDL binding and cholesteryl ester uptake. Using a blocking antibody against C323 region, we demonstrated that C323 is directly involved in HDL binding and likely an HDL binding site. Using C323G mutant transgenic mouse model, we further demonstrated that C323 of SR-BI is required for regulating plasma cholesterol levels in vivo. Using redox reagents, we showed that physiological relevant levels of H(2)O(2) upregulated the SR-BI-mediated cholesteryl ester uptake activity by 65%, whereas GSH or DTT significantly downregulated SR-BI-mediated cholesteryl ester uptake activity by 45%. C323 of SR-BI is critical for SR-BI-mediated HDL binding and cholesteryl ester uptake, and changes in redox status may be a regulatory factor modulating SR-BI-mediated cholesterol transport.
authors.affiliation: Department of Pediatrics, University of Kentucky Medical School, Lexington, KY.
authors.forename: Ling
authors.initials: L
authors.lastname: Guo
authors.affiliation: Department of Pediatrics, University of Kentucky Medical School, Lexington, KY; Taian Central Hospital, Taian, Shandong, China.
authors.forename: Min
authors.initials: M
authors.lastname: Chen
authors.affiliation: Department of Pediatrics, University of Kentucky Medical School, Lexington, KY; Taian Central Hospital, Taian, Shandong, China.
authors.forename: Zhiqing
authors.initials: Z
authors.lastname: Song
authors.affiliation: Saha Cardiovascular Research Center, University of Kentucky Medical School, Lexington, KY and.
authors.forename: Alan
authors.initials: A
authors.lastname: Daugherty
authors.affiliation: Department of Pediatrics, University of Kentucky Medical School, Lexington, KY; Saha Cardiovascular Research Center, University of Kentucky Medical School, Lexington, KY and; Graduate Center for Nutritional Sciences, University of Kentucky Medical School, Lexington, KY; and. Electronic address: xli2@email.uky.edu.
authors.forename: Xiang-An
authors.initials: XA
authors.lastname: Li
mesh_terms: D000818:Animals; D016466:CHO Cells; D021941:Caveolae / Q000378:metabolism; D002788:Cholesterol Esters / Q000097:blood / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003545:Cysteine; D005260:Female; D006801:Humans; D008075:Lipoproteins, HDL / Q000378:metabolism*; D051379:Mice; D010084:Oxidation-Reduction; D011485:Protein Binding; D021381:Protein Transport; D051122:Scavenger Receptors, Class B / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D002788:Cholesterol Esters; D008075:Lipoproteins, HDL; C495124:SCARB1 protein, human; D051122:Scavenger Receptors, Class B; D003545:Cysteine