Abstract
Apolipoprotein B-100 (apo B-100) contains putative lipid-associating regions that are, in part, responsible for its overall structure in human plasma low-density lipoproteins. Some of these regions have been identified by reassembly of the total tryptic peptides of apo B-100 with bovine brain sphingomyelin, 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC) and dimyristoylphos-phatidylcholine (DPMC). Although more than 500 tryptic peptides are predicted from the known number of arginines and lysines in apo B-100, significant amounts of only 13 peptides spontaneously associate with all three phospholipids. These peptides share some structural characteristics, as predicted by several algorithms, that distinguish them from the water-soluble apolipoproteins. Most apolipoproteins associate with lipids via amphipathic helices and are highly helical in native and reassembled lipoproteins. Analysis of all apo B-100 lipophilic peptides by circular dichroism and by use of a predictive algorithm reveals no evidence of amphipathic helices. Although the predictive algorithm suggested that the lipophilic peptides of apo B-100 contain the sequence determinants for β-sheet, no spectroscopic evidence for this structure was found. We conclude that the lipophilic regions of apo B-100 liberated by trypsinolysis are highly hydrophobic, although their secondary structures do not fit any simple model.
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References
Anantharamaiah, G. M. (1986).Methods Enzymol. 128, 627–647.
Bartlett, G. R. (1959).J. Biol. Chem. 234, 466–468.
Cardin, A. D., and Jackson, R. L. (1986).Biochim. Biophys. Acta 877, 366–371.
Chang, J. Y., Bauer, D., and Wittmann-Liebold, B. (1978).FEBS Lett. 93, 204–214.
Chou, P., and Fasman, G. (1978).Ann. Rev. Biochem. 47, 251–276.
Eisenberg, D., Weiss, R., and Terwilliger, T. C. (1982).Nature 299, 371–374.
Goldstein, J. L., and Brown, M. S. (1983). InThe Metabolic Basis of Inherited Disease (Stanbury, J. B.,et al., eds.), McGraw-Hill, New York, pp. 672–712.
Gotto, Jr., A. M., Levy, R. I., and Frederickson, D. S. (1968).Proc. Nat. Acad. Sci. (USA)60, 1436–1441.
Greenfield, N., and Fasman, G. D. (1969).Biochemistry 8, 4108–4116.
Havel, R. J., Goldstein, J. L., and Brown, M. S. (1980). InMetabolic Control and Disease (Bondy, P. K., and Rosenberg, L. E., eds.), Saunders, Philadelphia, pp. 393–494.
Hirose, N., Krivanek, M. A., Blankenship, D. T., Jackson, R. L., and Cardin, A. D. (1986).Arteriosclerosis 6, 529a.
Kaiser, E. T., and Kezdy, F. J. (1984).Science 223, 249–255.
Kane, J. P. (1983).Ann. Rev. Physiol. 45, 637–650.
Knott, T. J., Pease, R. J., Powell, L. M., Wallis, S. C., Rall, S. C., Innerarity, T. L., Blackhart, B., Taylor, W. H., Marcel, Y., Milne, R., Johnson, D., Fuller, M., Lusis, A. J., McCarthy, B. J., Mahley, R. W., Levy-Wilson, B., and Scott, J. (1986).Nature 323, 734–738.
Krebs, K. E., and Phillips, M. C. (1983).Biochim. Biophys. Acta 754, 227–230.
Krebs, K. E., and Phillips, M. C. (1984).FEBS Lett. 175, 263–266.
Kyte, J., and Doolittle, R. F. (1982).J. Mol. Biol. 154, 105–132.
Lux, S. E., Hirz, R., Shrager, R. I., and Gotto, Jr., A. M. (1972).J. Biol. Chem. 247, 2598–2606.
Mahley, R. W., Innerarity, T. L., Rall, S. C., and Weisgraber, K. H. (1984).J. Lipid Res. 25, 1277–1294.
Markwell, M-A. K., Haas, S. M., Bieber, L. L., and Tolbert, N. E. (1978).Anal. Biochem. 87, 206–210.
Osterman, D., Mora, R., Kezdy, F. J., Kaiser, E. T., and Meredith, S. C. (1984).J. Am. Chem. Soc. 106, 6845–6847.
Pownall, H. J., Knapp, R. D., Gotto, Jr., A. M., and Massey, J. B.FEBS Lett. 159, 17–23.
Pownall, H. J., Gotto, Jr., A. M., Knapp, R. D., and Massey, J. B. (1986).Biochem. Biophys. Res. Commun. 139, 202–208.
Pownall, H. J., Massey, J. B., Sparrow, J. T. and Gotto, Jr., A. M. (1987). InPlasma Lipoproteins (Gotto, A. M., ed.), Elsevier Scientific Publishers, Amsterdam, pp. 95–127.
Segrest, J. P., Jackson, R. L., Morrisett, J. D., and Gotto, Jr., A. M. (1974).FEBS Lett. 38, 247–253.
Weisgraber, K. H., Rall, S. C. Scott, J., Knott, T. J., and Mahley, R. W. (1986).Arteriosclerosis 6, 528a.
Yang, C. Y. (1979).Hoppe-Seyler's Z. Physiol. Chem. 360, 1673–1675.
Yang, C. Y., and Wakil, S. J. (1984).Anal. Biochem. 137, 54–57.
Yang, C. Y., Chen, S. H., Gianturco, S. H., Bradley, W. A., Sparrow, J. T., Tanimura, M., Li, W. H., Sparrow, D. A., DeLoof, H., Rosseneu, M., Lee, F. S., Gu, Z. W., Gotto, Jr., A. M., and Chan, L. (1986).Nature 323, 738–742.
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Yang, Cy., Kim, T.W., Pao, Q. et al. Structure and conformational analysis of lipid-associating peptides of apolipoprotein B-100 produced by trypsinolysis. J Protein Chem 8, 689–699 (1989). https://doi.org/10.1007/BF01024895
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DOI: https://doi.org/10.1007/BF01024895