Transfer of cholesteryl ester into high density lipoprotein by cholesteryl ester transfer protein: effect of HDL lipid and apoprotein content.
Recombinant high density lipoprotein (rHDL) particles were prepared by cosonication of purified lipids and human apoproteins and incubated with partly purified cholesteryl ester transfer protein (CETP) and low density lipoprotein (LDL) containing [3H]cholesteryl ester. Increasing the triglyceride content relative to cholesteryl ester in rHDL significantly decreased the ability of the particles to accept cholesteryl esters transferred by CETP. Kinetic analysis of the data was performed to numerically define the maximum velocity of lipid transfer, Tmax, and the HDL concentration required for half maximal velocity, KH. Increases in rHDL-triglyceride content were shown to result in a significant reduction in the Tmax without a major change in KH. When the free cholesterol content was increased relative to phospholipid, the ability of the particles to accept cholesteryl esters was also decreased in a similar manner. Conversely, rHDL prepared from purified apoprotein A-I, A-II, or mixtures of both, had significantly elevated Tmax and KH values for their interaction with CETP. The results suggest that increases in triglyceride or free cholesterol content of an rHDL particle decrease the catalytic ability of CETP by noncompetitive inhibition. In addition, some component(s) of HDL apoproteins, other than A-I or A-II, were shown to uncompetitively inhibit the activity of CETP, by modifying both Tmax and the KH for the reaction. This study has shown that altered HDL composition may have marked effects on the transfer and equilibration of cholesteryl esters within the HDL pool.
Sparks
D L
DL
Department of Pathology, University of British Columbia, Vancouver, Canada.
Pritchard
P H
PH
D001059:Apoproteins / Q000378:metabolism; D002352:Carrier Proteins / Q000302:isolation & purification / Q000502:physiology*; D002784:Cholesterol / Q000378:metabolism; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters / Q000378:metabolism*; D006023:Glycoproteins; D006801:Humans; D050356:Lipid Metabolism; D008075:Lipoproteins, HDL / Q000378:metabolism* / Q000648:ultrastructure; D010316:Particle Size; D011994:Recombinant Proteins; D014280:Triglycerides / Q000378:metabolism
D001059:Apoproteins; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters; D006023:Glycoproteins; D008075:Lipoproteins, HDL; D011994:Recombinant Proteins; D014280:Triglycerides; D002784:Cholesterol