Growth-promoting activity in serum-free medium of kallikreinlike arginylesteropeptidases from rat submaxillary gland.
The characterization and purification of the growth-promoting activity present in rat submaxillary gland extracts, known to be required for the proliferation of adipose precursor cells in serum-free medium, have been undertaken. Fractionation of the extracts by ion-exchange chromatography, gel filtration, and affinity chromatography on immobilized benzamidine allowed the copurification of the mitogenic activity with two distinct arginylesteropeptidases of apparent molecular weight 25,000; one of these enzymes has been purified to homogeneity and shown to be immunologically related to tonin, a well-characterized kallikreinlike protease from submaxillary gland. The specificity of both enzymes was similar to that of plasma and glandular kallikreins, as indicated by the relative rates of hydrolysis of peptide p-nitroanilide substrates. Prior treatment of the kallikreinlike proteases with phenylmethylsulfonylfluoride or aprotinin abolished completely both mitogenic and arginylesteropeptidase activities, indicating that enzymatic activity was essential for the manifestation of their growth-promoting ability. The kallikreinlike proteases from rat submaxillary gland were able to replace thrombin to support the proliferation of Chinese hamster lung fibroblasts in serum-free medium. These results underline the role of proteases in controlling cell growth and are discussed in light of adipose tissue development.
D000818:Animals; D002268:Carboxypeptidases / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003470:Culture Media; D006133:Growth Substances / Q000494:pharmacology*; D007610:Kallikreins; D007700:Kinetics; D001121:Lysine Carboxypeptidase / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D013363:Submandibular Gland / Q000201:enzymology*
D003470:Culture Media; D006133:Growth Substances; D011480:Protease Inhibitors; D002268:Carboxypeptidases; D001121:Lysine Carboxypeptidase; D007610:Kallikreins