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Kinetic Analysis of Lecithin:Cholesterol Acyltransferase Activity Toward Discoidal HDL

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  • Published:
Lipids

Abstract

The kinetics of lecithin:cholesterol acyltransferase(LCAT, EC 2.3.1.43)-catalyzed generation of cholesteryl ester in discoidal high density lipoproteins (HDL) was analyzed in terms of initial binding of LCAT to the disc surface followed by a three-state reaction of the hydrolysis of phosphatidylcholine sn-2 ester bond and acyl-enzyme formation. Cholesterol was considered as alcoholic nucleophile that increases the solvolysis rate of acyl-LCAT. The raw kinetic data of Sparks et al. (J Biol Chem 270:5151–5157, 1995) for four preparations of reconstituted discoidal HDL with a constant level of apolipoprotein A-I and palmitoyloleoylphosphatidylcholine per disc but with cholesterol in a lipid phase continuously increasing from 2.1 to 12.5 mol%, were analyzed in terms of the kinetic equation and a complete set of rate constants was obtained. Data at high cholesterol content do not indicate a saturation phenomenon, thus giving no evidence for a binding of cholesterol to the enzyme. This analysis may be used in the study of LCAT activation by exchangeable apolipoproteins and contribution of the HDL structure.

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Fig. 1

Abbreviations

apoA-I:

Apolipoprotein A-I

CE:

Cholesteryl ester

Chol:

Cholesterol

FFA:

Free fatty acid

HDL:

High density lipoproteins

LCAT:

Lecithin:cholesterol acyltransferase

LysoPtdCho:

Lysophosphatidylcholine

PtdCho:

Phosphatidylcholine

POPC:

1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine

RCT:

Reverse cholesterol transport

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Acknowledgments

The financial support of the Russian Foundation for Basic Research (grant # 10-04-00270) is gratefully acknowledged.

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  1. National Research Centre for Preventive Medicine, 10, Petroverigsky Street, 101990, Moscow, Russia

    Alexander D. Dergunov

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  1. Alexander D. Dergunov
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Correspondence to Alexander D. Dergunov.

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Dergunov, A.D. Kinetic Analysis of Lecithin:Cholesterol Acyltransferase Activity Toward Discoidal HDL. Lipids 46, 1075–1079 (2011). https://doi.org/10.1007/s11745-011-3613-6

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