ABCA1 and amphipathic apolipoproteins form high-affinity molecular complexes required for cholesterol efflux

M. L. Fitzgerald, A. L. Morris, A. Chroni, A. J. Mendez, V. I. Zannis, M. W. Freeman

Research output: Contribution to journalArticle

117 Scopus citations

Abstract

Apolipoproteins, such as apolipoprotein A-I (apoA-I), can stimulate cholesterol efflux from cells expressing the ATP binding cassette transporter A1 (ABCA1). The nature of the molecular interaction between these cholesterol acceptors and ABCA1 is controversial, and models suggesting a direct protein-protein interaction or indirect association have been proposed. To explore this issue, we performed competition binding and chemical cross-linking assays using six amphipathic plasma proteins and an 18 amino acid amphipathic helical peptide. All seven proteins stimulated lipid efflux and inhibited the cross-linking of apoA-I to ABCA1. Cross-linking of apoA-I to ABCA1 was saturable and occurred at high affinity (Kd of 7.0 ± 1.9 nM), as was cross-linking of apoA-II. After binding to ABCA1, apoA-I rapidly dissociated (half-life of 25 min) from the complex and was released back into the medium. A mutant form of ABCA1 (W590S) that avidly binds apoA-I but fails to promote cholesterol efflux released apoA-I with similar kinetics but without transfer of cholesterol to apoA-I. Thus, a high-affinity, saturable, protein-protein interaction occurs between ABCA1 and all of its amphipathic protein ligands. Dissociation of the complex leads to the cellular release of cholesterol and the apolipoprotein. However, dissociation is not dependent on cholesterol transfer, which is a clearly separable event, distinguishable by ABCA1 mutants.

Original languageEnglish (US)
Pages (from-to)287-294
Number of pages8
JournalJournal of Lipid Research
Volume45
Issue number2
DOIs
StatePublished - Feb 2004

Keywords

  • ATP binding cassette transporter A1
  • High density lipoprotein
  • Tangier Disease

ASJC Scopus subject areas

  • Endocrinology

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