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Measurements of Proteoglycan—Lipoprotein Interaction by Gel Mobility Shift Assay

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Lipoprotein Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 110))

Abstract

Proteoglycans (PGs) of the arterial intima form complexes with apo B-100 containing lipoproteins. Several lines of evidences indicate that this process IS a key pathogenic event for the development of atherosclerosis (for review, see ref. 1). First, retained apo B in early as well as advanced lesions is closely associated with arterial PGs (2, 3). Second, purified arterial PGs, partrcularly those from lesion-prone sites, bmd low-density lipoprotein (LDL) in vitro (4). Third, LDL from patrents with coronary-artery disease has a higher affinity for arterial PGs than LDL from apparently healthy subjects (5).

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References

  1. Hurt-Camejo, E, Olsson, U., Wiklund, O, Bondjers, G, and CameJo, G (1997) Cellular consequences of the association of apoB lipoproteins with proteoglycans Arterioscler. Thromb Vast Blof 17, 101l–1017.

    Google Scholar 

  2. Ylă-Herttuala, S., Solakivi, T., Hirvonen, J., et al. (1987) Glycosammoglycans and apolipoproteins B and A-I in human aortas. chemical and immunological analysis of lesion-free aortas from children and adults. Arteriosclerosis 7, 333–340

    PubMed  Google Scholar 

  3. Nievelstein-Post, P., Mottino, G., Fogelman, A, and Frank, J (1994) An ultrastructural study of lipoprotein accumulation in cardiac valves of the rabbit Arterioscler. Thromb 14, 115l–1161.

    Google Scholar 

  4. Cardoso, L. and Mourao, P. (1994) Glycosaminoglycan fractions from human arteries presenting diverse susceptibility to atherosclerosis have different binding affinities to plasma LDL. Arterioscler Thromb Vasc Biol 14, 115–124.

    CAS  Google Scholar 

  5. Camejo, G., Acquatella, H., and LaLaguna, F. (1980) The interaction of low density lipoprotein with arterial proteoglycans: an additional risk factor? Atherosclerosis 36, 55–65.

    Article  PubMed  CAS  Google Scholar 

  6. Camejo, G, Lòpez, A, Lòpez, F., and Quioñes, J. (1985) Interaction of low density lipoproteins with arterial proteoglycans the role of charge and sialic acid content. Atherosclerosis 55, 93–105.

    Article  PubMed  CAS  Google Scholar 

  7. Anber, V., Miller, J S., McConnell, M, Shepherd, J., and Packard, J (1997) Interaction of very-low-density, intermediate-density, and low-density hpoproteins with human arterial wall proteoglycans. Arterioscler Thromb. Vasc Biol 17, 2507–2514

    PubMed  CAS  Google Scholar 

  8. Wagner, W. D, Salisbury, B. G., and Rowe, H A. (1988) A proposed structure of chondronin 6-sulfate proteoglycans of human normal adjacent atherosclerotic plaque. Arterlosclerosis 6, 407–417

    Google Scholar 

  9. Jackson, R, Busch, S., and Cardin, A. (1991) Glycosaminoglycans.molecular properties, protein Interactions and role in physiological processes Physiolog. Rev 71, 481–539

    CAS  Google Scholar 

  10. Volker, W, Schmidt, A., and Buddecke, E (1987) Mapping of proteoglycans in human arterial tissue. Eur J Ceil. Biol 45, 72–79.

    CAS  Google Scholar 

  11. Wight, T N (1996) The vascular extracellular matrix, in Atherosclerosis and Coronary Artery Dlseuse (Fuster, V., Ross, R, and Topol, E. J, eds.), Philadelphia: Lippmcott-Raven, pp 42l–440

    Google Scholar 

  12. Camejo, G, Olofsson, S-O., Lopez, F, and Bondjers, G. (1989) Identification of apoB-100 segments mediating the interaction of low density lipoproteins with arterial proteoglycans Arterlosclerosis 8, 368–377.

    Google Scholar 

  13. Hurt-Camejo, E. and CameJo, G. (1997) Potential involvement of type II phospholipase A2 in atherosclerosis. Atherosclerosis 132, 1–8.

    Article  PubMed  CAS  Google Scholar 

  14. Olsson, U., Camejo, G., and Bondjers, G. (1993) Binding of a synthetic apohpoprotein B-100 peptide and peptide analogues to chondroitm-6-sulfate effect of the lipid environment. Biochemistry 32, 1858–1865.

    Article  PubMed  CAS  Google Scholar 

  15. Linden, T., Bondjers, G., Carrrejo, G., Bergstrand, R., Wilhelmsen, L, and Wiklund, O (1989) Affimty of LDL to a human arterial proteoglycan among male survivors of myocardial infarction. Eur J Clan Invest 19, 38–44

    CAS  Google Scholar 

  16. Anber, V., Griffin, B., McConnell, M., Packard, C, and Sheperd, J (1996) Influence of plasma lipid and LDL-subfraction profile on the interaction between low density lipoprotem with human arterial wall proteoglycans. Atherosclerosis 124, 261–271.

    Article  PubMed  CAS  Google Scholar 

  17. Wiklund, O., Bondjers, G., Wright, I., and Camejo, G. (1996) Insoluble complex formation between LDL and arterial proteoglycans in relation to serum lipid levels and effect of lipid lowering drugs. Atherosclerosis 119, 57–68

    Article  PubMed  CAS  Google Scholar 

  18. Havel, R, Eder, H, and Bragdon, J H. (1995) The distribution and chemical composition of ultracentrifugally separated lipoprotems in human serum J Clin Invest 34, 1345–1353

    Article  Google Scholar 

  19. Camejo, G, Fager, G, Rosengren, B. E. H and Bondjers, G (1993) Binding of low density lipoprotems by proteoglycans synthesized by proliferating and qutescent human arterial smooth muscle cells. J. Biol Chem 268, 14,13l–14,137.

    Google Scholar 

  20. Sartipy, P., Johansen, B., Cannejo, G., Rosengren, B., Bondjers, G, and Hurt-Camejo, E (1996) Binding of human phosphohpase A2 type II to proteoglycans, differential effect of glycosammoglycans on enzyme activity. J. Biol Chem 271, 26,307–26,314

    Article  PubMed  CAS  Google Scholar 

  21. Goldberg, R. and Kolibas, L. (1990) An improved method for determining proteoglycans synthesized by chondrocytes in culture Connect Tissue Res 24, 265–275

    Article  PubMed  CAS  Google Scholar 

  22. Revzin, A. (1989) Gel electrophoresis assays for DNA-protein mteractrons BloTechnlques 7, 346–355.

    CAS  Google Scholar 

  23. Epps, D. E., Raub, T. J., and Kezdy, F. J. (1995) A general, wide-range spectrofluorometric method for measuring the site-specific affinities of drugs toward human serum albumin Anal Blochem. 227, 342–350.

    Article  CAS  Google Scholar 

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© 1998 Humana Press Inc., Totowa, NJ

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Hurt-Camejo, E., Camejo, G., Sartipy, P. (1998). Measurements of Proteoglycan—Lipoprotein Interaction by Gel Mobility Shift Assay. In: Ordovas, J.M. (eds) Lipoprotein Protocols. Methods in Molecular Biology™, vol 110. Humana Press. https://doi.org/10.1385/1-59259-582-0:267

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  • DOI: https://doi.org/10.1385/1-59259-582-0:267

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-420-4

  • Online ISBN: 978-1-59259-582-2

  • eBook Packages: Springer Protocols

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