Reaction of lecithin: cholesterol acyltransferase with micellar substrates. Effect of particle sizes.
Micellar, discoidal complexes were prepared from L-alpha-dipalmitoylphosphatidylcholine (DPPC) or egg phosphatidylcholine (egg-PC), cholesterol, and human apolipoprotein A-I by the cholate dialysis method. Reaction mixtures containing from 70:7:1 to 500:50:1, PC/cholesterol/apolipoprotein A-I (mol/mol) were fractionated by gel-filtration into various complex fractions. The isolated DPPC complexes ranged in size from 103 to 380 A in diameter, and in composition from 70:7:1 to 470:45:1, PC/cholesterol/apolipoprotein A-I (mol/mol), respectively. In contrast, the isolated egg-PC complexes only ranged in size from 105 to 214 A in diameter, and in composition from 65:5:1 to 153:17:1, PC/cholesterol/apolipoprotein A-I (mol/mol), respectively. Measurements of fluorescence wavelength maxima and fluorescence polarization of tryptophan residues of apolipoprotein A-I, in both series of complexes, revealed uniform spectral properties for all the egg-PC containing complexes. The DPPC complexes, on the other hand, had maxima in the fluorescence parameters for complexes with diameters around 200 A. When reacted with purified human lecithin:cholesterol acyltransferase, either at constant apolipoprotein A-I or at constant lipid concentration, all egg-PC complexes had very similar reaction rates, but the DPPC complex series exhibited major differences in reactivity. Minima in reaction rates occurred for DPPC complexes around 200 A in diameter, and optimal rates were observed with the small discoidal complexes (110 A in diameter). These reaction rates correlate well with the apolipoprotein A-I fluorescence properties and indicate that the apolipoprotein structure, reflected at the interface with phosphatidylcholine, may be the most important factor in determining complex reactivity with lecithin:cholesterol acyltransferase.
D016632:Apolipoprotein A-I; D001053:Apolipoproteins / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism; D003102:Colloids; D005454:Fluorescence Polarization; D006801:Humans; D008823:Micelles; D008854:Microscopy, Electron; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000378:metabolism*; D010713:Phosphatidylcholines; D011663:Pulmonary Surfactants
D016632:Apolipoprotein A-I; D001053:Apolipoproteins; D003102:Colloids; D008823:Micelles; D010713:Phosphatidylcholines; D011663:Pulmonary Surfactants; D002784:Cholesterol; D007862:Phosphatidylcholine-Sterol O-Acyltransferase