Purification and partial characterization of a carboxypeptidase from the limpet (Patella vulgata) |
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Authors: | G M Hass |
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Affiliation: | Department of Bacteriology and Biochemistry, University of Idaho, Moscow, Idaho 83843 USA |
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Abstract: | A carboxypeptidase A-like enzyme has been purified to apparent homogeneity from commercially available acetone powder from the visceral hump of the limpet, Patella vulgata. A two-step procedure involving affinity chromatography on ?-amino-N-caproyl-d-phenylalanine-Sepharose and gel filtration resulted in a 3000-fold purification with an 80% yield. The enzyme is a single polypeptide chain of Mr = 40,000 and exhibits both peptidase and esterase activities, which are characterized by dramatic excess substrate inhibition. Inhibition studies suggest that a metal ion is required for activity and demonstrate that the affinity label, N-bromoacetyl-N-methyl-l-phenylalanine, and a polypeptide carboxypeptidase inhibitor from potatoes (apparent Ki approx. 2 nm) are effective against the limpet enzyme. |
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