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Journal of Lipid Research, Vol. 44, 1132-1142, June 2003
Copyright © 2003 by Lipid Research, Inc.

A quantitative analysis of apolipoprotein binding to SR-BI

Stephen T. Thuahnai^*, Sissel Lund-Katz^*, G. M. Anantharamaiah, David L. Williams and Michael C. Phillips^1,*

^* Division of GI/Nutrition, Lipid Research Group, The Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-4318
Biochemistry and Molecular Genetics, Atherosclerosis Research Unit, University of Alabama at Birmingham, Birmingham, AL 35294
Department of Pharmacological Sciences, University Medical Center, State University of New York at Stony Brook, Stony Brook, NY 11794

¹ To whom correspondence should be addressed. e-mail: phillipsmi{at}email.chop.edu

Competitive binding experiments were performed using Y1-BS1 adrenal cells to provide information about the interaction of HDL apolipoproteins with scavenger receptor class B, type I (SR-BI). Exchangeable apolipoproteins apolipoprotein A-I (apoA-I), apoA-II, apoE-2, apoE-3, and apoE-4 as phospholipid complexes bind like HDL₃ to SR-BI via their multiple amphipathic -helices; the concentrations required to reduce the binding of HDL₃ to SR-BI by 50% (IC₅₀) were similar and in the range of 35–50 µg protein/ml. In the case of apoA-I, peptides corresponding to segments 1–85, 44–65, 44–87, 149–243, and 209–241 all had the same IC₅₀ as each other (P = 0.86), showing that a specific amino acid sequence in apoA-I is not responsible for the interaction with SR-BI. The distribution of charged residues in the amphipathic -helix affects the interaction, with class A and Y helices binding better than class G* helices. Synthetic -helical peptides composed of either L or D amino acids can bind equally to the receptor. Association with phospholipid increases the amount of apolipoprotein binding to SR-BI without altering the affinity of binding. Lipid-free apolipoproteins compete only partially with the binding of HDL to SR-BI, whereas lipidated apolipoproteins compete fully.

These results are consistent with the existence of more than one type of apolipoprotein binding site on SR-BI.

Abbreviations: CE, cholesteryl ester; CNBr, cyanogen bromide; DMPC, 1,2-dimyristoyl phosphatidylcholine; GdnHCl, guanidine hydrochloride; MLV, multilamellar vesicle; POPC, 1-palmitoyl-2-oleoyl phosphatidylcholine; rHDL, reconstituted HDL; SR-BI, scavenger receptor class B, type I; SUV, small unilamellar vesicle

Supplementary key words scavenger receptor class B, type I • lipoprotein metabolism • high density lipoprotein • lipoprotein receptor • amphipathic -helix

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