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Journal of Lipid Research, Vol. 23, 97-104, January 1982
Copyright © 1982 by Lipid Research, Inc.
Donner Laboratory, Lawrence Berkeley Laboratory, University of California, Berkeley CA 94720
Density gradient ultracentrifugation of low density lipoproteins (LDL) from 12 normal subjects showed multiple, distinct isopycnic bands. Densitometric scanning of the gradient tubes revealed that each band could be assigned to one of four density intervals and that the boundaries of these intervals were consistent among all the subjects. Analytic ultracentrifuge flotation (Sf0) rates were assigned to the four density intervals, and there was a strong correlation between peak Sf0 rate and peak isopycnic banding position (Rf) of the LDL in the 12 subjects. The Sf0 value corresponding to the boundary between the two most buoyant LDL density subgroups was 7.5. This value is close to that previously demonstrated to define two LDL subdivisions (Sf0 0-7 and Sf0 7-12) that were discriminated by differing concentrations in men and women, and differing statistical relationships with levels of HDL and VLDL in a normal population. Further delineation of distinct subspecies of LDL was afforded by electrophoresis in 2-16% gradient polyacrylamide gels. Densitometric scans of proteinstained gels revealed multiple peaks, and particle diameters were assigned to these peaks using calibration markers. Particles of diameter ≥ 280 Å included both IDL and Lp(a), the latter defined by pre-ßbeta; mobility on agarose electrophoresis and density > 1.050 g/ml. LDL particles with diameters 220-272 Å could be grouped into seven size intervals defined by modes in the distribution of gradient gel electrophoretic peaks in LDL from a group of 68 healthy men and women. Particle diameters of the major peaks in each of seven density subfractions decreased with increasing density of the fractions. However, particles within each of the size groups were distributed across a range of densities. Use of a lipid-staining procedure allowed identification of electrophoretic bands in whole plasma which corresponded to those seen in isolated LDL, eliminating the possibility that ultracentrifugation was responsible for formation of the subspecies detected by the gradient gel procedure. The application of density gradient ultracentrifugation and gradient gel electrophoresis provides a means of characterizing LDL from normal humans in terms of multiple distinct subpopulations which may also prove to have differing metabolic and pathologic properties.—Krauss, R. M., and D. J. Burke. Identification of multiple subclasses of plasma low density lipoproteins in normal humans.
Supplementary key words serum lipoproteins • very low density lipoproteins • high density lipoproteins • density gradient ultracentrifugation • analytic ultracentrifugation • polyacrylamide gel electrophoresis
Submitted on February 13, 1981
Revised on July 20, 1981
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