Two-dimensional CNBr peptide patterns of collagen types I, II and III

CNBr peptides from the alpha 1-chains of collagen types I, II and III have been fractionated by a high resolution two-dimensional mapping procedure. Radioactive collagen chains were synthesized without deamination of lysyl residues by rabbit cells in culture and were treated with pepsin and purified prior to cleavage by CNBr. The resultant peptides from each collagen type were separately fractionated by a two-dimensional mapping procedure consisting of non-equilibrium isoelectric focusing in the first dimension and sodium dodecyl sulfate electrophoresis in the second dimension. Unique peptide maps were obtained for each type of alpha-chain and only small amounts of sample were required. This method will be useful for unambiquous identification of isolated collagen chains, identification of collagens in complex mixtures, and the rapid identification of cross-linked peptides.