A [4,5-3H]lysine:[14C]lysine dual-label method to measure lysine hydroxylation in collagen

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 noncollagen 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 measureme..