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Cholesterol Determines and Limits rHDL Formation From Human Plasma Apolipoprotein A-II and Phospholipid Membranes

Cholesterol Determines and Limits rHDL Formation From Human Plasma Apolipoprotein A-II and Phospholipid Membranes

G Randall Bassett, Baiba K Gillard, Henry J Pownall

Biochemistry. 2012 Oct 30;51(43):8627-35.

PMID: 23025327

Abstract:

Apolipoprotein (apo) A-II, the second most abundant protein after apo A-I of human plasma high-density lipoproteins (HDL), is the most lipophilic of the exchangeable apolipoproteins. The rate of microsolubilization of dimyristoylphosphatidylcholine (DMPC) membranes by apo A-I to give rHDL increases as the level of membrane free cholesterol (FC) increases up to 20 mol % when the level of reaction decreases to nil. Given its greater lipophilicity, we tested the hypothesis that apo A-II and its reduced and carboxymethylated monomer (rcm apo A-II) would form rHDL at a membrane FC content of >20 mol %. According to turbidimetric titrations, the DMPC/apo A-II stoichiometry is 65/1 (moles to moles). At this stoichiometry, apo A-II forms rHDL from DMPC and FC. Contrary to our hypothesis, apo A-II, like apo A-I, reacts poorly with DMPC containing ≥20 mol % FC. The rate of formation of rHDL from rcm apo A-II and DMPC at all FC mole percentages is faster than that of apo A-II but nil at 20 mol % FC. In parallel reactions, monomeric and dimeric apo A-II form large FC-rich rHDL coexisting with smaller FC-poor rHDL; increasing the FC mole percentage increases the number and size of FC-rich rHDL. On the basis of the compositions of coexisting large and small rHDL, the free energy of transfer of FC from the smallest to the largest particle is approximately -1.2 kJ. On the basis of our data, we propose a model in which apo A-I and apo A-II bind to DMPC via surface defects that disappear at 20 mol % FC. These data suggest apo A-II-containing HDL formed intrahepatically are likely cholesterol-rich compared to the smaller intracellular lipid-poor apo A-I HDL.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR4247070	Apolipoprotein A-II from human plasma			Price

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