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Journal of Lipid Research, Vol 29, 1557-1571, Copyright © 1988 by Lipid Research, Inc.
RW James, D Hochstrasser, JD Tissot, M Funk, R Appel, F Barja, C Pellegrini, AF Muller and D Pometta
The protein heterogeneity of fractions isolated by immunoaffinity
chromatography on anti-apolipoprotein A-I and anti-apolipoprotein A-II
affinity columns was analyzed by high resolution two-dimensional gel
electrophoresis. The two-dimensional gel electrophoresis profiles of the
fractions were analyzed and automatically compared by the computer system
MELANIE. Fractions containing apolipoproteins A-I + A-II and only A-I as
the major protein components have been isolated from plasma and from high
density lipoproteins prepared by ultracentrifugation. Similarities between
the profiles of the fractions, as indicated by two- dimensional gel
electrophoresis, suggested that those derived from plasma were equivalent
to those from high density lipoproteins (HDL), which are particulate in
nature. The established apolipoproteins (A-I, A-II, A-IV, C, D, and E) were
visible and enriched in fractions from both plasma and HDL. However,
plasma-derived fractions showed a much greater degree of protein
heterogeneity due largely to enrichment in bands corresponding to six
additional proteins. They were present in trace amounts in fractions
isolated from HDL and certain of the proteins were visible in
two-dimensional gel electrophoresis profiles of the plasma. These proteins
are considered to be specifically associated with the
immunoaffinity-isolated particles. They have been characterized in terms of
Mr and pI. Computer-assisted measurements of protein spot-staining
intensities suggest an asymmetric distribution of the proteins (as well as
the established apolipoproteins), with four showing greater prominence in
particles containing apolipoprotein A-I but no apolipoprotein A-II.
ARTICLES
Protein heterogeneity of lipoprotein particles containing apolipoprotein A-I without apolipoprotein A-II and apolipoprotein A-I with apolipoprotein A-II isolated from human plasma
Division de Diabetologie, Hopital Cantonal Universitaire, Geneva, Switzerland.
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