Role of apolipoprotein E in hepatic lipase catalyzed hydrolysis of phospholipid in high-density lipoproteins.
We reported earlier that hepatic lipase (HL)-catalyzed hydrolysis of phospholipid monolayers is activated by apolipoprotein (apo) E [Thuren et al. (1991b) J. Biol. Chem. 266, 4853-4861]. On the basis of these studies, it was postulated that apoE-rich high-density lipoproteins (HDL) were preferred substrates for HL. In the present study, we tested this hypothesis, as well as further characterizing the activation of HL hydrolysis of phospholipid by apoE. The apoE-rich HDL, referred to as HDL-I, were isolated by heparin-Sepharose chromatography, and the phospholipid hydrolysis by HL was compared to an apoE-poor HDL, designated HDL-II. The hydrolysis of HDL-I phosphatidylcholine was approximately 3-fold higher than HDL-II, supporting the hypothesis that HL preferably hydrolyzes the phospholipids in apoE-rich HDL. In order to gain additional insight into the nature of the activation, we used phospholipid monolayers as model systems. Comparison of the ability of the two thrombolytic fragments of apoE (22 kDa, residues 1-191; 12 kDa, residues 192-299) revealed that only the 12-kDa fragment was capable of activating the hydrolysis of phospholipid by HL (1.75-fold). However, activation was less than with the intact protein (2.8-fold for apoE3), suggesting that the intact protein was required for full activation. The fact that the 12-kDa fragment, which represents a major lipid region of the protein, did activate HL suggests that activation occurs at the lipid-water interface.(ABSTRACT TRUNCATED AT 250 WORDS)
Thuren
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Department of Biochemistry, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27157-1016.
Weisgraber
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KH
Sisson
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P
Waite
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D000042:Absorption; D000328:Adult; D000818:Animals; D001057:Apolipoproteins E / Q000737:chemistry / Q000502:physiology*; D002384:Catalysis; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D006868:Hydrolysis / Q000187:drug effects; D007536:Isomerism; D008049:Lipase / Q000378:metabolism* / Q000502:physiology; D008075:Lipoproteins, HDL / Q000737:chemistry / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D008297:Male; D010743:Phospholipids / Q000378:metabolism*; D051381:Rats; D013379:Substrate Specificity
D001057:Apolipoproteins E; D008075:Lipoproteins, HDL; D010743:Phospholipids; D008049:Lipase