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Journal of Lipid Research, Vol. 10, 68-76, January 1969
Copyright © 1969 by Lipid Research, Inc.

Particle size and protein content of six fractions of the S_f > 20 plasma lipoproteins isolated by density gradient centrifugation

W. J. Lossow , F. T. Lindgren , J. C. Murchio , G. R. Stevens , and L. C. Jensen

Donner Laboratory, Lawrence Radiation Laboratory, University of California, Berkeley, California 94720, and Air and Industrial Hygiene Laboratory, Division of Laboratories, California State Department of Public Health, Berkeley, California 94704

A procedure is described for the separation of plasma S_f > 400 and S_f 20-400 lipoproteins each into three fractions. Serum samples are overlayered with a sodium chloride density gradient in a preparative ultracentrifuge tube and thin layers are removed at the top of the tube after successive centrifugations at different speeds in a swinging bucket rotor.

The procedure was evaluated by electron microscopy of the S_f > 400 lipoprotein fractions and schlieren analysis of the S_f 20-400 lipoprotein fractions. Protein content of each fraction was measured by elemental N, C, H, and lipid-P analysis. Protein coverage was calculated for all fractions on the assumption that there is a surface layer 20 A thick.

For the entire S_f > 400 lipoprotein spectrum and for a part of the S_f 20-400 lipoprotein distribution the proportion of surface covered by protein was constant (approximately 20% coverage). Therefore, for these portions of the lipoprotein spectrum, the increase in surface: volume ratio as particle size decreases is approximately compensated for by an increase in the weight percentage of protein.

Supplementary key words chylomicrons • very low density lipoproteins • electron microscopy • analytical centrifugation • protein coverage

Submitted on July 12, 1968
Accepted on September 17, 1968

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