The interaction of human apolipoprotein C-I with sub-micellar phospholipid

Benjamin W. Atcliffe, Christopher A. MacRaild, Paul R. Gooley, Geoffrey J. Howlett

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10 Citations (Scopus)

Abstract

Mature human apolipoprotein C-I (apoC-I), comprising 57 amino acids, is the smallest member of the plasma apolipoprotein family. Amphipathic helical regions within apoC-I, common to this class of proteins, are mediators of lipid binding, a process that underlies the functional properties of apoC-I, including the capacity to activate the plasma enzyme LCAT, to disrupt apoE mediated receptor interactions and to inhibit cholesterol ester transfer protein. To examine apoC-I/phospholipid interactions, we have developed an expression system in Escherichia coli to obtain purified apoC-I with yields of approximately 4-5 mg per L of culture. The purified product has properties similar to plasma-derived apoC-I including self-association in the lipid-free state and induced α-helical content in the presence of egg-yolk phosphatidylcholine and dimyristoylglycerophosphocholine vesicles. We chose the short-chain phospholipid, dihexanoylglycerophosphocholine (Hex2Gro-PCho), to examine the interaction of apoC-I with submicellar phospholipid. Circular dichroism spectroscopy and cross-linking experiments show that apoC-I acquires helical content and remains self-associated at submicellar concentrations of Hex2Gro-PCho (4 mM). Sedimentation equilibrium studies of apoC-I at submicellar levels of Hex2Gro-PCho and analysis of the effects of apoC-I on the 1H NMR spectrum of Hex2Gro-PCho indicate micelle induction by apoC-I, and establish the capacity of apoC-I to assemble individual phospholipid molecules.

Original languageEnglish
Pages (from-to)2838-2846
Number of pages9
JournalEuropean Journal of Biochemistry
Volume268
Issue number10
DOIs
Publication statusPublished - 27 Sep 2001
Externally publishedYes

Keywords

  • apoC-I
  • HexGro-PCho
  • Protein-lipid interaction
  • Sedimentation equilibrium
  • Submicellar phospholipid

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