A [4,5-3H]lysine:[14C]lysine dual-label method to measure lysine hydroxylation in collagen |
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| Authors: | M Chojkier W Holderman J F Bateman |
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| Institution: | 1. Department of Medicine, Veterans Administration Medical Center San Diego, California USA;2. University of California, San Diego, California USA;3. Department of Paediatrics, University of Melbourne, Royal Children''s Hospital, Parkville, Victoria 3052, Australia |
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| Abstract: | A new method has been developed to determine the extent of lysine hydroxylation in newly synthesized collagen. This method relies on the measurement of changes in the ratio of 3H]lysine:14C]lysine in collagenase digests, resulting from loss of tritium from the C-5 position of lysine during hydroxylation. Lysine hydroxylation can be measured in the presence of large amounts of noncollagen proteins, and simultaneous quantitation of the relative rates of collagen and non-collagen protein production is obtained. The dual-label lysine method is simple, rapid, and accurate. There was a very good correlation between this method and column chromatography procedures currently used for the measurement of lysine hydroxylation. |
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