A rapid and large-scale isolation of renin from mouse submaxillary gland by pepstatin-aminohexyl-agarose affinity chromatography.
A pressor enzyme, renin, was purified about 60-fold by chromatography on an affinity column including pepstatin-aminohexyl-agarose with a high yield of 83% from the homogenate of adult mouse submaxillary glands. The renin obtained by the one-step purification was electrophoretically homogenous on SDS-polyacrylamide gel and was as active as an absolutely pure renin. The renin  purified by the affinity column could be separated into five active components by chromatography on CM-cellulose. Each of these renins gave a symmetrical elution profile on the CM-cellulose column and a discrete protein band on polyacrylamide gel electrophoresis at pH 8.6. Administration of nanogram quantities of each of the two major renin fractions to nephrectomized rats caused a sustained rise of blood pressure and decrease in sensitivity of the animal to angiotensin II. This rapid and large-scale purification method using pepstatin-aminohexyl-agarose eliminates all four fractionation steps reported previously for the isolation of mouse submaxillary gland renin.
D000818:Animals; D001681:Biological Assay; D001794:Blood Pressure / Q000187:drug effects; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange; D008297:Male; D051379:Mice; D010436:Pepstatins; D051381:Rats; D012083:Renin / Q000302:isolation & purification* / Q000494:pharmacology; D012685:Sepharose / Q000031:analogs & derivatives; D013363:Submandibular Gland / Q000201:enzymology*
D010436:Pepstatins; C024903:aminohexyl-sepharose; D012685:Sepharose; D012083:Renin