Purification and partial characterization of a high molecular weight metalloproteinase from the venom of Crotalus adamanteus (eastern diamondback rattlesnake).

Chromatography of Crotalus adamanteus venom on CM-Sepharose, Cibacron Blue-Sepharose and Phenyl-Sepharose, followed by gel filtration on Ultrogel AcA 44, has resulted in the isolation in homogeneous condition of a metalloproteinase active on casein and hide powder azure. The proteinase has an alkaline isoelectric point, and the trivial name proteinase B ('basic proteinase') is suggested to distinguish it from previously characterized C. admanteus metalloproteinases. Proteinase B is a single chain glycoprotein containing one free sulfhydryl group and having a molecular weight of 60,000. Proteinase B was inactivated by treatment with EDTA, but exposure to phenylmethylsulfonyl fluoride had no effect on proteolytic activity. Proteinase B lacked hemorrhagic activity and did not digest chromogenic substrates specific for thrombin, plasmin or plasma kallikrein.