Two-step affinity-chromatographic purification of cathepsin D from pig myometrium with high yield.
Cathepsin D was purified by two-step affinity chromatography on concanavalin A-- and pepstatin--Sepharose. The main purification was achieved by washing the enzyme bound to the pepstatin--Sepharose column with buffered 6 M-urea. This step separated cathepsin D from all low- and high-molecular-weight impurities. Although the 1700-fold purified acid proteinase was homogeneous on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, it still showed microheterogeneity.
D000818:Animals; D002402:Cathepsin D; D002403:Cathepsins / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D003208:Concanavalin A; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D009215:Myometrium / Q000201:enzymology*; D010436:Pepstatins; D012685:Sepharose; D013552:Swine; D014508:Urea; D014599:Uterus / Q000201:enzymology*
D010436:Pepstatins; D003208:Concanavalin A; D014508:Urea; D012685:Sepharose; D002403:Cathepsins; D002402:Cathepsin D