Procollagen and collagen produced by normal bovine corneal stroma fibroblasts in cell culture.

Procollagen and collagen were isolated from the culture medium of normal bovine corneal stromal fibroblasts. DEAE-cellulose chromatography was used to separate the collagen molecules from the different procollagens present. One collagen and four procollagen peaks were isolated and biochemically characterized. All the procollagen fractions and the collagen fraction yielded, after limited pepsin or chymotrypsin digestion followed by CNBr digestion, molecules that correspond to (alpha 1)2 alpha 2 exclusively. Thus only type I collagen is found in the growth medium of of bovine corneal stromal fibroblast cultures. Each of the individual procollagen peaks contained pro-alpha chains having molecular weights of 120,000, 150,000, 165,000, 180,000, and 190,000 daltons, according to their elution position on DEAE-cellulose. The presence of type I procollagen molecules having pro-alpha chains of 165,000, 180,000, and 190,000 daltons has not previously been reported and probably represents higher-molecular-weight precursor intermediates. The amino acid compositions of the different procollagen fractions are unique, and each contains relatively large amounts of cysteine and tryptophan. Carbohydrate analysis, cyanogen bromide peptide analysis, electron microscopy of SLS-crystallities, and SDS-polyacrylamide gel electrophoresis were used to further characterize the procollagen and collagen molecules.