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pubmed:abstractText |
A method for the isolation and purification of pyridinoline from bone collagen was developed, with the use of sulphonated polystyrene resins. The analytical techniques were used to quantify pyridinoline, for which hydroxyallysine is a known precursor, in a wide range of tissues. The structure of pyridinoline proposed by Fujimoto, Moriguchi, Ishida & Hayashi [(1978) Biochem. Biophys. Res. Commun. 84, 52-57] was confirmed by 13C-n.m.r. spectroscopy and fast-atom-bombardment mass spectrometry. At concentrations greater than about 0.1 mM, pyridinoline exhibited altered fluorescence properties that were consistent with excimer formation. From alkali hydrolysates of several different tissues, a fluorescent compound was purified by gel filtration and ion-exchange chromatography and was shown to be galactosylpyridinoline. This derivative was very labile to acid treatment compared with the bifunctional cross-link analogues, and was completely converted into free pyridinoline by heating at 108 degrees C for 8 h in 0.1 M-HCl. Galactosylpyridinoline was also partially converted into free pyridinoline by prolonged alkali hydrolysis. This lability, which could also apply to other multifunctional cross-link derivatives, may explain the fact that no disaccharide derivatives of pyridinoline were isolated.
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