Synthesis of collagenous proteins by pulmonary type II epithelial cells.
We have investigated the production of collagenous proteins by primary cultures of rat lung epithelial cells (type II pneumocytes). Three major bacterial collagenase-sensitive chains were synthesized and secreted into the medium between 12 and 36 h of culture. Two of the chains comigrated on sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis (SDS-PAGE) with radiolabeled type IV procollagen (PC) chains isolated from adult rat lung (Mr = 185,000 and 170,000 after reduction) and were coprecipitated with monospecific antibodies to type IV collagen. Cyanogen bromide (CNBr) peptide maps of the chromatographically purified chains were identical to maps of rat lung type IV PC, and confirmed the identity of these chains as pro alpha 1(IV) and pro alpha 2(IV). Type IV PC was the major high molecular weight collagen in the cell layer, and a fraction of the newly synthesized type IV PC was selectively deposited on the substratum together with newly synthesized fibronectin. Type II cells also secreted a low molecular weight, non-disulfide-bonded, collagenase-sensitive protein (Mr = 19,000, collagen standards; Mr = 26,000, globular standards). The protein coeluted with type IV PC from DEAE-cellulose but was resolved from native type IV on CM-cellulose. The protein was not precipitated with polyclonal antibodies to type IV collagen or rat surfactant apoprotein. These studies further demonstrate the heterogeneity of collagenous macro-molecules synthesized by lung epithelial cells in vitro. We suggest that interactions between pneumocyte-derived fibronectin and type IV procollagen contribute to the formation of the epithelial basement membrane and to the attachment of these cells in normal or injured lung.
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006868:Hydrolysis; D007120:Immunochemistry; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D008297:Male; D010449:Peptide Mapping; D011347:Procollagen / Q000096:biosynthesis* / Q000302:isolation & purification; D051381:Rats; D011919:Rats, Inbred Strains
D011347:Procollagen