Collagenase digestion demonstrates carboxy-terminal crosslinking in acid-soluble collagen.

Among the products of the collagenase cleavage of Type I acid-soluble collagen from calf and rabbit tendons, there can be found fragments with the lengths of half alpha-chains. Because purified collagenase cleaves the alpha-chains three-quarters of the length from the amino-terminus, the presence of half-length chains is evidence for the occurrence of crosslinks between two carboxy-terminal, quarter-length fragments, The collagen preparations were reduced with [3H]borohydride, the collagenase-cleaved fragments were separated by gel electrophoresis, and their 3H-labeled crosslink derivatives were analyzed. The major labeled components in the half-length chains were the reduced aldol condensation product and hydroxylysinonorleucine. These experiments demonstrate that the carboxy-terminal telopeptides in monomer-enriched collagen samples form aldol crosslinks which are probably intramolecular, but some intermolecular aldol and aldimine crosslinks may also be formed.