Abstract
Lecithin–cholesterol acyltransferase (LCAT) is the major enzyme responsible for the esterification of free cholesterol on plasma lipoproteins, which is a key step in the reverse cholesterol transport pathway. The measurement of plasma LCAT activity not only is important in the diagnosis of patients with genetic or acquired LCAT deficiency but is also valuable in calculating cardiovascular risk, as well as in research studies of lipoprotein metabolism. In this chapter, we describe a convenient LCAT assay based on the use of an apoA-I mimetic peptide. The proteoliposome substrate used in this assay for LCAT is easily made with the peptide and can be stored by deep freezing without significant loss of activity.
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References
Jonas A (2000) Lecithin cholesterol acyltransferase. Biochim Biophys Acta 1529:245–256
Fielding CJ, Fielding PE (1995) Molecular physiology of reverse cholesterol transport. J Lipid Res 36:211–228
Calabresi L, Moleri E, Franceschini G (2006) LCAT deficiency: molecular genetics, lipid/lipoprotein phenotype and atherosclerosis. Future Lipidol 1:241–245
Hoeg JM, Santamarina-Fojo S, Berard AM, Cornhill JF, Herderick EE, Feldman S, Haudenschild CC, Vaisman BL, Hoyt RF, Demosky SJ Jr, Kauffman RD, Hazel CM, Marcovina S, Brewer HB Jr (1996) Overexpression of lecithin cholesterol acyltransferase in transgenic rabbits prevents diet-induced atherosclerosis. Proc Natl Acad Sci U S A 93:11448–11453
Sakai N, Vaisman BL, Koch CA, Hoyt RF Jr, Meyn SM, Talley GD, Paiz JA, Brewer HB Jr, Santamarina-Fojo S (1997) Targeted disruption of the mouse lecithin:cholesterol acyltransferase (LCAT) gene. Generation of a new animal model for human LCAT deficiency. J Biol Chem 272:7506–7510
National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) (2002) Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) final report. Circulation 106:3143–3421
Gotto AM Jr, Brinton EA (2004) Assessing low levels of high-density lipoprotein cholesterol as a risk factor in coronary heart disease: a working group report and update. J Am Coll Cardiol 43:717–724
Feig JE, Shamir R, Fisher EA (2008) Atheroprotective effects of HDL: beyond reverse cholesterol transport. Curr Drug Targets 9:196–203
Bots ML, Visseren FL, Evans GW, Riley WA, Revkin JH, Tegeler CH, Shear CL, Duggan WT, Vicari RM, Grobbee DE, Kastelein JJ, RADIANCE 2 Investigators (2007) Torcetrapib and carotid intima-media thickness in mixed dyslipidaemia (RADIANCE 2 study): a randomised, double-blind trial. Lancet 370:153–160
Frohlich J, Dobiasova M (2003) Fractional esterification rate of cholesterol and ratio of triglycerides to HDL-cholesterol are powerful predictors of positive findings on coronary angiography. Clin Chem 49:1873–1880
Chen C-H, Albers JJ (1982) Characterization of proteoliposomes containing apoprotein A-I: a new substrate for the measurement of lecithin: cholesterol acyltransferase activity. J Lipid Res 23:680–691
Albers JJ, Chen CH, Lacko AG (1986) Isolation, characterization, and assay of lecithin-cholesterol acyltransferase. Methods Enzymol 129:763–783
Gillett MP, Owen JS (1992) Cholesterol esterifying enzyme—lecithin: cholesterol acyltransferase (LCAT) and acylcoenzyme A:cholesterol acyltransferase (ACAT). In: Converse CA, Skinner ER (eds) Lipoprotein analysis. A practical approach. IRL, Oxford, pp 187–201
Dassuex J-L, Sekul R, Bittner K, Cornut I, Metz G, Dufourcq J (1999) Apolipoprotein A-I agonists and their use to treat dyslipidemic disorders. US Patent 6,004,925
Sethi AA, Amar M, Shamburek RD, Remaley AT (2007) Apolipoprotein AI mimetic peptides: possible new agents for the treatment of atherosclerosis. Curr Opin Investig Drugs 8:201–212
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Vaisman, B.L., Remaley, A.T. (2013). Measurement of Lecithin–Cholesterol Acyltransferase Activity with the Use of a Peptide-Proteoliposome Substrate. In: Freeman, L. (eds) Lipoproteins and Cardiovascular Disease. Methods in Molecular Biology, vol 1027. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-369-5_16
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DOI: https://doi.org/10.1007/978-1-60327-369-5_16
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-60327-369-5
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