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

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

Research output: Contribution to journalArticleResearchpeer-review

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

Cite this

Atcliffe, Benjamin W. ; MacRaild, Christopher A. ; Gooley, Paul R. ; Howlett, Geoffrey J. / The interaction of human apolipoprotein C-I with sub-micellar phospholipid. In: European Journal of Biochemistry. 2001 ; Vol. 268, No. 10. pp. 2838-2846.
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The interaction of human apolipoprotein C-I with sub-micellar phospholipid. / Atcliffe, Benjamin W.; MacRaild, Christopher A.; Gooley, Paul R.; Howlett, Geoffrey J.

In: European Journal of Biochemistry, Vol. 268, No. 10, 27.09.2001, p. 2838-2846.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - The interaction of human apolipoprotein C-I with sub-micellar phospholipid

AU - Atcliffe, Benjamin W.

AU - MacRaild, Christopher A.

AU - Gooley, Paul R.

AU - Howlett, Geoffrey J.

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AB - 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.

KW - apoC-I

KW - HexGro-PCho

KW - Protein-lipid interaction

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JF - European Journal of Biochemistry

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