title: Collagen biosynthesis by cultured Chinese hamster lung cells. Cell-free synthesis of procollagen alpha chains.
abstract: Cell-free extracts from the HTl clone of cultured Chinese hamster lung cells efficiently promote the incorporation of proline into newly synthesized material, 50% of which is digestible to small peptides by highly purified bacterial collagenase. The synthesis of the these products occurs under optimal protein synthesis conditions and is inhibited by puromycin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the cell-free synthesized material reveals a major collagenase sensitive peak (20% of the total product) at mol wt 165 000 which is reflected by a collagenase sensitive material of similar size in the culture medium. Two additional collagenase digestible species (mol wt 95000 and 65000), each having a corresponding secreted product, are generated by the cell-free system. These results are consistent with the concept that procollagen is formed by the association of three individually translated pro alphachains. The data further constitute the report of a highly active homologous cell-free system capable of pro alpha chain biosynthesis derived from a cultured cell line that is a practical source for pro alphachain biosynthesis derived from a cultured cell line that is a practical source for proalpha chain mRNA as well as a unique system for elucidating regulatory mechanisms involved in collagen biosynthesis.
mesh_terms: D002460:Cell Line; D002474:Cell-Free System; D007930:Leucine / Q000378:metabolism; D046911:Macromolecular Substances; D003012:Microbial Collagenase / Q000378:metabolism; D008970:Molecular Weight; D011347:Procollagen / Q000032:analysis / Q000096:biosynthesis*; D011392:Proline / Q000378:metabolism
chemical_list: D046911:Macromolecular Substances; D011347:Procollagen; D011392:Proline; D003012:Microbial Collagenase; D007930:Leucine