Quantitation of type I, III, and V collagens in human tissue samples by high-performance liquid chromatography of selected cyanogen bromide peptides.

A method to determine the proportions of the major fiber-forming collagens (types I, III, and V) in noncartilaginous human tissues is presented. The procedure relies on direct solubilization of tissue collagen as cyanogen bromide peptides. The peptides are subjected to cation exchange chromatography followed by gel permeation chromatography in a manner consistent with the rapid resolution and quantitation of relatively low-molecular-weight marker peptides for each collagen.

Immunolocalization of types V and XI collagen in cartilage using monoclonal antibodies.

Monoclonal antibodies produced against pepsin-solubilized newborn rat skin type V collagen [alpha 1(V)]2 alpha 2(V), and chondrosarcoma type XI collagen [alpha 1(XI) alpha 2(XI) alpha 3(XI)] are used to localize the collagens in sections of the chondrosarcoma as well as the normal rat knee joint by indirect immunofluorescence. Immunostaining for type V collagen shows strong cellular staining of chondrocytes; while the interstitial matrix as well as the lacunae are not stained. In contrast, antitype XI stains not only chondrocytes, but the extracellular compartments as well.

Isolation and characterization of the cyanogen bromide peptides of the collagen 2 alpha chain from a transplantable rat chondrosarcoma.

The rat collagen 2 alpha chain was isolated from a transplantable Swarm chondrosarcoma following limited pepsin proteolysis. The chromatographically purified 2 alpha chain when cleaved with cyanogen bromide yields nine peptides which have been isolated and characterized with respect to their molecular weight and their amino acid composition. Eight of these peptides can be rapidly separated by gel-permeation high-performance liquid chromatography and retrieved for further purification by ion-exchange chromatography.