Detection and isolation of inactive, large molecular weight renin in human kidney and plasma.
Estimation of apparent molecular weight (mw) of inactive renin by gel filtration of human plasma was found to be inaccurate when "acid activation" or "cryoactivation" was used for detection; recoveries were only 5% to 20%. Trypsin activation produced greater recoveries, but the apparent elution volume of inactive renin varied with the concentration of trypsin used; the presence of trypsin inhibitors increased trypsin requirements to 100 to 200 micrograms/ml in the 60,000 dalton region, while low protein concentration in the 50,000 dalton region resulted in destruction of renin by as little as 10 microgram/ml trypsin. A composite trypsin-activated inactive renin peak corresponded to a mw of 56,000 +/- 1500 daltons (104% to 120% recovery), while active plasma renin was 48,000 +/- 2000 daltons. When this prorenin-like substance was isolated by affinity chromatography, it was found to be completely inactive. It was also nearly free of trypsin inhibitors, so that a single trypsin concentration correctly identified and confirmed the elution characteristics of inactive renin peak following gel filtration. The apparent mw of trypsin-activated inactive renin was slightly lower (52,000 daltons) than that of inactive renin. Human renal cortex was also found to contain a trypsin-activable form of renin. Like plasma inactive renin, it could be isolated by chromatography on Cibacron blue-agarose (Affi-Gel blue). It was found to be completely inactive following passage over a pepstatin affinity column. This inactive renal renin, as well as a similar substance in perfusate of normal human kidney, had a mw of 49,500 +/- 1000, while active renal renin was 39,500 +/- 500. Trypsin-activated inactive renal renin had a mw of 46,500 +/- 500; its pH optimum was identical with that of active renal renin, and it no longer bound to Cibacron blue-agarose. We conclude that both human plasma and kidney contain an inactive, prorenin-like substance that can be detected reliably by trypsin activation. There appear to be slight differences in the apparent mw of plasma renins and kidney renin, but the similarity of other characteristics suggests that the inactive, prorenin-like substances in renal cortex, renal perfusate, and plasma may be one and the same substance.
- Copyright © 1981 by American Heart Association