Discrimination of collagen types by methionine-specific stain

Abstract

Chloroglycyl-L-methioninatoplatinum (II) (Pt-GLM) specifically labels methionine residues in collagen. Rat Type I and calf Type III collagen segment long spacing aggregates show differences in Pt-GLM bands, in agreement with known methionine positions. Reconstituted fibers formed from the same two collagens again demonstrate some differences in band pattern, which can be understood in terms of the amino acid sequences and the known packing of molecules in the fiber. Native collagen fibers have been extracted from rat and chick tendon (predominantly Type I collagen). These have been stained with Pt-GLM and studied in the Johns Hopkins STEM and a CTEM. Density profiles along the collagen fibers have been obtained, and those for successive 670-A repeats averaged, by computer programs. The position of peaks in the average density profiles for these various fibers show very good correlation with the known locations of Met residues. The band patterns and density profiles for native and reconstituted Type I fibers are very similar and differ from those of reconstituted Type III fibers.