Biosynthesis and in vitro translation of type IV procollagens

The present paper describes how epitheilial cells, cultured from bovine anterior lens capsule explants, synthesize and secrete procollagen type IV polypeptide chains α1(IV). and α2(IV). Metabolic labeling of these cells with [C]proline for different time intervals and subsequent analysis by SDS/polyacrylamide gel electrophoresis revealed the presence of two polypeptide chains with apparent molecular masses of 180 kDa and 170 kDa. The procollagens were bacterial‐collagenase‐sensitive and were specifically immunoprecipitated by antibodies raised against the 7S domain of type IV collagen. Type IV procollagen poly(A)‐rich RNA was was isolated from cultured lens capsule cells and translated in a reticulocyte lysate cell‐free system. Two polypetptides with apparent molecular lens capsule cells and translated in a reticulocyte lysate cell‐free system. Two polypeptides with apparent molecular masses of 152 kDa and 145 kDa were identified as procollagen type IV unmodified chains by gel electrophoresis, collagenase digestion and specific immunoprecipitation. During experiments in which cells were labeled in the presence of α,α′‐bipyridyl, type IV procollagen appeared as one major band comigrating with a 145 kDa polypeptide on SDS‐gel electrophoresis. Copyright © 1985, Wiley Blackwell. All rights reserved