Abstract
The primary functional role of collagen is as a supporting tissue and it is now well established that the aggregated forms of the collagen monomers are stabilized to provide mechanical strength by a series of intermolecular crosslinks. These links are formed by oxidative deamination of the ε-amino group of the single lysine in the amino and carboxy-telopeptides by lysyl oxidase. The aldehyde thus formed reacts with an ε-amino group of a lysine at a specific point in the triple helix because of the quarter-staggered end-overlap alignment of the molecules in the fibers. The chemistry of these crosslinks is dependent on both the nature and age of the collagenous tissue (1,2). Differences in the crosslinks are because of the degree of hydroxylation of both the telopeptide and the specific lysine in the triple helix. Thus, the amounts of intermediate crosslinks present in immature tissue, dehydro-hydroxylysinonorleucine (A-HLNL), and hydroxylysino-keto-norleucine (HLKNL) may vary considerably between tissues, e.g., rat tail tendon and skin contain A-HLNL whereas cartilage and bone contain predominantly HLKNL.
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References
Bailey, A. J., Light, N. D., and Atkins, E. D. T. (1980) Chemical cross-linking restrictions on models for the molecular organization of the collagen fibre. Nature 288, 408–410.
Knott, L. and Bailey, A. J. (1998) Collagen cross-links in mineralising tissues: A review of their chemistry, function and clinical relevance. Bone 22, 181–187.
Yamauchi, M., London, R. E., Guemat, C., Hashimoto, F., and Mechanic, G. L. (1987) Structure and function of a stable histidine-based tri-functional cross-link in skin collagen. J. Biol. Chem. 262, 11,428–11,434.
Eyre, D. R. and Oguchi, H. (1980) The hydroxypyridinium cross-link of skeletal collagen. Biochem. Biophys. Res. Commun. 92, 403–410.
Light, N. D. and Bailey, A. J. (1982) Covalent cross-links in collagen, in Methods in Enzymology, vol. 82A, pp. 360–372.
Sims, T. J. and Bailey, A. J. (1992) Quantitative analysis of collagen and elastin cross-links using a single-column system. J. Chromatog. 582, 49–55.
Robins, S. P. (1982) Analysis of the cross-linking components in collagen and elastin. Methods Biochem. Anal. 28, 329–379.
Scott, J. E., Hughes, E. W., and Shuttleworth, A. (1981) A collagen associated Ehrlich chromogen: A pyrrollic cross-link? Biosci. Rep. 209, 263–264.
Paul, R. G. and Bailey, A. J. (1996) Glycation of collagen. The basis of its central role in the late complications of ageing and diabetes. Interntl. J. Biochem. Cell Biol. 28, 1297–1310.
Partridge, S. M. (1970) Isolation and characterisation of elastin, in Chemistry and Molecular Biology of the Intercellular Matrix, vol. 1, Academic, New York, pp. 593–616
Avery, N. C. and Bailey, A. J. (1995) An efficient method for the isolation of intramuscular collagen. Meat Sci. 41, 97–100.
Grant, R. A. (1964) Application of the auto-analyser to connective tissue analysis. J. Clin.Pathol. 17, 685–691.
Riley, G., Harrall, R. L., Constant, C. R. Chard, M. D., Cawston, T. E., and Hazleman, B. L. (1994) Tendon degeneration and chronic shoulder pain: changes in the collagen composition of the human rotator cuff tendons in rotator cuff tendinitis. Ann. Rheum. Dis. 53, 359–366.
Sell, D. R. and Monnier, V. M. (1989) Structure elucidation of a senescence cross-link from human extracellular matrix-Implication of pentoses in the ageing pro-cess. J. Biol. Chem. 264, 21,597–21,602.
Dyer, D. G., Blackledge, A., Thorpe, S. R., and Baynes, J. W. (1991) Formation of pentosidine during non-enzymatic browning of proteins by glucose. J. Biol. Chem. 268, 11,654–11,660.
Takahashi, M., Ohishi, T., Aoshima, H., Kushida, K, Inoue T., and Horiuchi, K. (1993) Prefractionation with cation exchanger for determination of intermolecu-lar cross-links, pyridinoline and pentosidine, in hydrolysates. J. Liq. Chrom. 16, 1355–1370.
Avery, N. C. (1996) The use of solid phase cartridges as a pre-fractionation step in the quantitation of intermolecular collagen cross-links and advanced glycation end-products. J. Liq. Chrom. 19, 1831–1848.
Avery, N. C. and Light, N. D. (1985) Re-packing reversed-phase high perfor-mance liquid chromatography columns as a means of regenerating column effi-ciency and prolonging packing life. J. Chrom. 328, 347–352.
Dawson, R. M. C., Elliott, D. C., Elliott, W. H., and Jones, K. M. (1986) Data for Biochemical Research. 3rd Ed. Clarendon, Oxford.
Bailey, A. J., Sims, T. J., Avery, N. C., and Halligan, E. P. (1995) Non-enzymic glycation of fibrous collagen; reaction products of glucose and ribose. Biochem. J. 305, 385–390.
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Sims, T.J., Avery, N.C., Bailey, A.J. (2000). Quantitative Determination of Collagen Crosslinks. In: Streuli, C.H., Grant, M.E. (eds) Extracellular Matrix Protocols. Methods in Molecular Biology™, vol 139. Humana Press. https://doi.org/10.1385/1-59259-063-2:11
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DOI: https://doi.org/10.1385/1-59259-063-2:11
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