Demonstration of renin activity in purified rat Leydig cells: evidence for the existence of an endogenous inactive (latent) form of enzyme.
Previous histochemical studies demonstrated the specific localization of immunoreactive renin-like substance in Leydig cells of rat testes. The studies reported herein demonstrate a specific renin enzyme activity in the two purified populations of Leydig cells (I and II) of mature rat testes. Leydig cells of both populations also exhibited an inactive (latent) renin which was activated by the sulfhydryl reagents dithiothreitol, beta-mercaptoethanol, glutathione, and cysteine but not by limited proteolysis by trypsin, which is a characteristic activating agent for prorenin or inactive renin of the zymogen type. The activation of latent renin by dithiothreitol produced approximately 5-and 10-fold increases in renin activity in Leydig cell populations I and II, respectively. Active and latent renin showed strong affinity to an antirat renin immunoglobulin-Sepharose column, indicating a close immunological relationship of latent renin to active renin. Both active and latent renin from Leydig cell populations (I and II) exhibited the pH optimum of 6.0. The gel filtration of Leydig cell extracts characteristically revealed that the apparent mol wts of active and latent renin were 39,000 and 48,000, respectively. Both active and latent renin in Leydig cells remained almost at similar levels through four continuous subcultures. The activity of latent renin slightly increased during the four consecutive subcultures, while active renin levels remained almost constant.
D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D004789:Enzyme Activation; D007700:Kinetics; D007985:Leydig Cells / Q000166:cytology / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D012083:Renin / Q000302:isolation & purification / Q000378:metabolism*; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology
D013439:Sulfhydryl Reagents; D014357:Trypsin; D012083:Renin