Enhanced synthesis and secretion of type IV collagen and entactin during adipose conversion of 3T3-L1 cells and production of unorthodox laminin complex.
Synthesis and secretion of basement membrane proteins by 3T3-L1 adipocytes were studied by metabolic labeling of the cells with [35S]methionine. Enhanced synthesis and secretion of many polypeptides of high molecular weight were observed by stimulating the adipose conversion of 3T3-L1 fibroblasts with dexamethasone, 1-methyl-3-isobutylxanthine, and insulin. Among these polypeptides, alpha 1(IV) and alpha 2(IV) chains of collagen were identified based on specific immunoprecipitation and digestion with bacterial collagenase. Synthesis and secretion of alpha 1(IV) and alpha 2(IV) chains were negligible in the fibroblasts, but remarkably enhanced in adipocytes. Based on specific immunoprecipitation of a sulfated polypeptide of 150 kDa, enhanced (6-fold) synthesis and secretion of entactin were also demonstrated. Immunoprecipitation with anti-laminin antiserum showed synthesis of three polypeptides with sizes corresponding to B subunits but failed to demonstrate synthesis of the A subunit. Synthesis of these laminin-related polypeptides remained constant during the conversion. Nonreducing sodium dodecyl sulfate electrophoresis showed intracellular assembly of three laminin-related polypeptides into binary and ternary complexes in a similar sequence of AB1B2 formation via B1B2 in embryonal carcinoma F9 (Morita, A., Sugimoto, E., and Kitagawa, Y. (1985) Biochem. J. 229, 259-264). The ternary complex of laminin in 3T3-L1 cells had a size significantly smaller than AB1B2 complex in F9 cells. In this complex, a novel subunit appears to take the place of the A subunit. Thus, a novel laminin complex is produced by 3T3-L1 cells.
Aratani
Y
Y
Institute for Biochemical Regulation, School of Agriculture, Nagoya University, Japan.
Kitagawa
Y
Y
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism; D000818:Animals; D001485:Basement Membrane / Q000187:drug effects / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis*; D003907:Dexamethasone / Q000494:pharmacology; D006023:Glycoproteins / Q000096:biosynthesis*; D007328:Insulin / Q000494:pharmacology; D007797:Laminin / Q000096:biosynthesis*; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D008715:Methionine / Q000378:metabolism; D051379:Mice
D006023:Glycoproteins; D007328:Insulin; D007797:Laminin; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; C029967:nidogen; D003907:Dexamethasone; D003094:Collagen; D008715:Methionine; D015056:1-Methyl-3-isobutylxanthine