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Journal of Lipid Research, Vol. 18, 734-744, November 1977
Copyright © 1977 by Lipid Research, Inc.

Separation and characterization of plasma lipoproteins of rhesus monkeys (Macaca mulatta)

Lawrence L. Rudel , Dianne G. Greene , and Ramesh Shah

Arteriosclerosis Research Center, Bowman Gray School of Medicine of Wake Forest University, Winston-Salem, NC 27103

A group of 14 adult male rhesus monkeys was maintained on a low cholesterol-high fat diet. Periodically, animals were fasted and blood samples were taken for characterization of the plasma lipoproteins. Complete separation of individual plasma lipoprotein classes was not achieved by traditional sequential ultracentrifugation techniques. Rather, initial separation of lipoprotein classes according to size was effected and density centrifugation was used subsequently for further separation. At least six lipoprotein fractions were identified, each of which was unique as defined by the properties of size, density (d), and electrophoretic mobility. These lipoprotein fractions were characterized by determination of chemical compositions and apoprotein patterns. The lipoproteins present in highest concentration in these monkeys were designated as region IV lipoproteins. This fraction had -migration on agarose electrophoresis, 1.063 < d < 1.225, and the size, composition, and apoprotein pattern characteristic of HDL. No fewer than three fractions were identified with densities that overlapped the 1.019 < d < 1.063 range. Of these, the fraction designated as region III lipoproteins was present in highest concentration, had ßbeta;-migration by agarose electrophoresis, a predominant B apoprotein, and a chemical composition and size characteristic of LDL. Two larger subfractions, identified as region II lipoproteins, were separated from each other at a density of 1.050 g/ml. Agarose electrophoresis showed that the fraction with d < 1.050 had a migration intermediate between ßbeta; and pre-ßbeta;. The chemical composition and apoprotein pattern were consistent with the possibility that these lipoproteins were remnants of VLDL catabolism. The fraction with d > 1.050, had pre-ßbeta; mobility and a size and composition similar to the Lp(a) lipoprotein in plasma of human beings. At least two VLDL subfractions, identified as region I and II_a lipoproteins, were found although both were present in very low concentrations. Region I lipoproteins were larger and contained relatively more cholesteryl ester and more of the apoproteins that migrated with the mobility of apo-B and arg-rich apoprotein in SDS-polyacrylamide gel electrophoresis. Some of the region I lipoproteins were ßbeta;-migrating by agarose electrophoresis. These results suggested the possibility that a ßbeta;-migrating VLDL was present in these normal animals.

Supplementary key words agarose gel chromatography • agarose electrophoresis • apoproteins • cholesterol • cholesteryl esters • high density lipoproteins • low density lipoproteins • polyacrylamide gel electrophoresis • very low density lipoproteins

Submitted on February 3, 1977
Accepted on May 11, 1977

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