Abstract
A method for the quantitative assessment of apolipoprotein distributions among baboon serum lipoproteins is described. The method combines the precise and reproducible separation of lipoproteins by polyacrylamide gradient gel electrophoresis with the specificity of immunoblotting. The method permits the measurement of distributions for any apolipoprotein for which there are antibodies available. Radioactive secondary antibodies are used to expose X-ray film, and distributions are determined by densitometry. Absorbance is linearly related to both antigen and antibody concentrations. The method is reproducible, with the mean coefficient of variation calculated to be 0.118, and has a high repeatability (r 2=0.97). The immunoblotting method can be employed to measure the fine details of lipoprotein phenotypes as they are influenced by genotype and environment.
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References
Aitchison, J. (1986).The Statistical Analysis of Compositional Data Chapman and Hall, New York.
Babiak, J., Gong, E. L., Nichols, A. V., Forte, T. M., Kuehl, T. J., and McGill, H. C., Jr. (1984). Characterization of HDL and lipoproteins intermediate to LDL and HDL in the serum of pedigreed baboons fed an atherogenic diet.Atherosclerosis 5227.
Bisgaier, C. L., Sachdev, O. P., Megna, L., and Glickman, R. M. (1985). Distribution of apolipoprotein A-IV in human plasma.J. Lipid Res. 2611.
Breslow, J. L. (1991). Lipoprotein transport gene abnormalities underlying coronary heart disease susceptibility.Annu. Rev. Med. 42357.
Chapman, M. J. (1986). Comparative analysis of mammalian plasma lipoproteins.Meth. Enzymol. 12870.
Cheng, M.-L., Woodford, S. C., Hilburn, J. L., and VandeBerg, J. L. (1986). A novel system for storage of sera frozen in small aliquots.J. Biochem. Biophys. Methods 1347.
Cheng, M.-L., Kammerer, C. M., Lowe, W. F., Dyke, B., and VandeBerg, J. L. (1988). Method for quantitating cholesterol in subfractions of serum lipoproteins separated by gradient gel electrophoresis.Biochem. Genet. 26657.
Fainaru, M., Havel, R. J., and Imaizumi, K. (1977). Apoprotein content of plasma lipoproteins of the rat separated by gel chromatography or ultracentrifugation.Biochem. Med. 17347.
Green, P. H. R., Glickman, R. M., Riley, J. W., and Quinet, E. (1980). Human apolipoprotein A-IV. Intestinal origin and distribution in plasma.J. Clin. Invest. 65911.
Greenwood, F. C., Hunter, W. M., and Glover, J. S. (1963). The preparation of131I-labelled human growth hormone of high specific radioactivity.Biochem. J. 89114.
Hancock, K., and Tsang, V. C. W. (1983). India ink staining of proteins on nitrocellulose paper.Anal. Biochem. 133157.
Hixson, J. E., Borenstein, S., Cox, L. A., Rainwater, D. L., and VandeBerg, J. L. (1988). The baboon gene for apolipoprotein A-I: Characterization of a cDNA clone and identification of DNA polymorphisms for genetic studies of cholesterol metabolism.Gene 74483.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227680.
Lefevre, M., Goudy-Lefevre, J. C., and Roheim, P. S. (1987). Gradient gel electrophoresis-immunoblot analysis (GGEI): A sensitive method for apolipoprotein profile determinations.J. Lipid Res. 281495.
Lefevre, M., Sloop, C. H., and Roheim, P. S. (1988). Characterization of dog prenodal peripheral lymph lipoproteins. Evidence for the peripheral formation of lipoprotein-unassociated apoA-I with slow pre-β electrophoretic mobility.J. Lipid Res. 291139.
Mahley, R. W., Innerarity, T. L., Rall, S. C., Jr., and Weisgraber, K. H. (1984). Plasma lipoproteins: Apolipoprotein structure and function.J. Lipid Res. 251277.
McGill, H. C., Jr., McMahan, C. A., Kushwaha, R. S., Mott, G. E., and Carey, K. D. (1986). Dietary effects on serum lipoproteins of dyslipoproteinemic baboons with high HDL1.Arteriosclerosis 6651.
Morrison, D. (1976).Multivariate Statistical Methods 2nd ed., McGraw-Hill, New York.
Musliner, T. A., Michenfelder, H. J., and Krauss, R. M. (1988). Interactions of high density lipoproteins with very low and low density lipoproteins during lipolysis.J. Lipid Res. 29349.
Neter, J., and Wasserman, W. (1974).Applied Linear Statistical Models Richard D. Irwin, Homewood, IL.
Rainwater, D. L., Blangero, J., Hixson, J. E., Mott, G. E., and VandeBerg, J. L. (1991). Lecithin:cholesterol acyltransferase (LCAT) polymorphism influences high-density lipoprotein (HDL) composition in baboons.FASEB J. 5:A1287.
Towbin, H., Staehelin, T., and Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications.Proc. Natl. Acad. Sci. USA 764350.
VandeBerg, J. L., and Cheng, M.-L. (1986). The genetics of baboons in biomedical research. In Else, J. G., and Lee, P. C. (eds.),Primate Evolution Cambridge University Press, Cambridge, p. 317.
Vezina, C. A., Milne, R. W., Weech, P. K., and Marcel, Y. L. (1988). Apolipoprotein distribution in human lipoproteins separated by polyacrylamide gradient gel electrophoresis.J. Lipid Res. 29573.
Williams-Blangero, S., VandeBerg, J. L., Blangero, J., Konigsberg, L., and Dyke, B. (1990). Genetic differentiation between baboon subspecies: Relevance for biomedical research.Am. J. Primatol. 2067.
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This work was supported in part by grant HL28972 and Contract No. HV53030 from the National Institutes of Health.
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Rainwater, D.L., Kammerer, C.M., Cheng, ML. et al. Distribution of specific apolipoproteins determined by immunoblotting of baboon lipoproteins resolved by polyacrylamide gradient gel electrophoresis. Biochem Genet 30, 143–158 (1992). https://doi.org/10.1007/BF02399705
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DOI: https://doi.org/10.1007/BF02399705