Human spleen cysteineproteinase inhibitor. Purification, fractionation into isoelectric variants and some properties of the variants.
The papain inhibitor from human spleen was purified by extraction in isotonic sucrose, acetone fractionation, papain-Sepharose affinity chromatography and gel filtration on Sephadex G-50. The purified inhibitor was fractionated by electrofocusing into four major isoelectric variants with pI values of 4.7, 5.0, 6.0 and 6.5. These variants can be classified into two groups: the acidic type, comprising the variants with pI 4.7 and 5.0, and the neutral type, comprising the variants with pI 6.0 and 6.5. The following properties distinguish the two types: 1. Immunological properties: antibodies raised against either of the neutral variants precipitated both of these, but not the acidic variants. The antiserum against the human epidermal cysteineproteinase inhibitor precipitated the acidic variants, but not the neutral variants. 2. Molecular size: two-dimensional electrophoresis of the purified inhibitor gave molecular weights of 11400 for the acidic variants and 12000 for the neutral variants. The pI 6.0 variant contained two compounds with molecular weights of 12000 and 12800. 3. Enzyme spectrum: human cathepsin B was inhibited by the acidic type, while the neutral type was a poor inhibitor. Both types inhibited cathepsin H, papain, ficin and bromelain, although the inhibition of bromelain did not exceed 70%. Human cathepsin D, bovine trypsin and chymotrypsin and porcine elastase were not inhibited by either type.
D000368:Aged; D003546:Cysteine Endopeptidases; D004355:Drug Stability; D004817:Epidermis / Q000201:enzymology; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007525:Isoelectric Focusing; D008875:Middle Aged; D008970:Molecular Weight; D010206:Papain / Q000037:antagonists & inhibitors; D011480:Protease Inhibitors / Q000276:immunology / Q000302:isolation & purification*; D013154:Spleen / Q000201:enzymology*
D000939:Epitopes; D011480:Protease Inhibitors; D003546:Cysteine Endopeptidases; D010206:Papain