A [4,5-3H]lysine:[14C]lysine dual-label method to measure lysine hydroxylation in collagen.
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.
D000818:Animals; D002250:Carbon Radioisotopes; D002642:Chick Embryo; D003094:Collagen / Q000032:analysis*; D006900:Hydroxylation; D006901:Hydroxylysine / Q000032:analysis*; D011506:Proteins / Q000032:analysis; D014316:Tritium
D002250:Carbon Radioisotopes; D011506:Proteins; D014316:Tritium; D006901:Hydroxylysine; D003094:Collagen