Properties of discoidal complexes of human apolipoprotein A-I with phosphatidylcholines containing various fatty acid chains.
In this study we demonstrate that apolipoprotein A-I determined the common size classes of discoidal particles formed with numerous phosphatidylcholines, and with ether analogs of phosphatidylcholines. We show furthermore, that the nature of the lipids dictates the distribution of particles among the different size classes. These experiments were performed with discoidal complexes containing various phospholipids (phosphatidylcholines with saturated and unsaturated fatty acid chains of different lengths and the ether analog of 1-palmitoyl-2-oleoylphosphatidylcholine), cholesterol, and human apolipoprotein A-I, prepared by the sodium cholate dialysis method, and fractionated by Bio-Gel A-5m gel-filtration chromatography. The complex preparations were analyzed in terms of their average composition, spectral properties of the apolipoprotein, and the dynamic behavior of the lipid domains. Nondenaturing gradient gel electrophoresis was used to analyze the size classes of particles present in the complex preparations. Starting with reaction mixtures containing around 100:1, phospholipid/apolipoprotein A-I molar ratios, complexes were isolated with molar ratios from 40:1 to 100:1. In most complexes apolipoprotein A-I had high levels of alpha-helical structure (65-77% alpha-helix), and tryptophan residues in a nonpolar environment. The lipid domains of complexes exhibited the dynamic behavior expected of the main phospholipid components. In the average size range from 90 to 100 A diameters, discrete particle classes with 80, 87, 102, 108, or 112 A Stokes diameters were observed for all the complexes containing different phospholipids. These discrete, recurring particle sizes are attributed to distinct apolipoprotein A-I conformations and variable lipid content.
D016632:Apolipoprotein A-I; D001054:Apolipoproteins A / Q000032:analysis*; D002784:Cholesterol / Q000032:analysis; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003720:Densitometry; D004586:Electrophoresis / Q000379:methods; D005227:Fatty Acids / Q000032:analysis*; D005231:Fatty Acids, Unsaturated / Q000032:analysis; D005454:Fluorescence Polarization; D006801:Humans; D008823:Micelles; D010316:Particle Size; D010713:Phosphatidylcholines / Q000032:analysis*
D016632:Apolipoprotein A-I; D001054:Apolipoproteins A; D005227:Fatty Acids; D005231:Fatty Acids, Unsaturated; D008823:Micelles; D010713:Phosphatidylcholines; D002784:Cholesterol