Zinc-mediated thermal stabilization of carboxypeptidase A.
In this study we investigated the contribution of Zn ions to the catalytic and structural thermostability of carboxypeptidase A (CPA). Structural studies on CPA molecule, performed in the presence of a number of ligands, demonstrated the multiple binding models around Zn ions which may affect the enzyme functions. Zinc was reported to bind at various sites in the CPA molecule at room temperature leading to inhibition of its enzymic activity. In this study we found that binding of Zn to CPA molecule followed by exposure to 50 degrees C did not inhibit the enzymic activity but activates and protects it against heat denaturation. The stabilization effect was found to be dependent on the increasing Zn/CPA ratios. The moderate changes of CPA activity as well as the UV and fluorescence spectra analyses indicate that the main function of the newly introduced zinc atoms is structural rather than catalytical.
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Department of Molecular Microbiology and Biotechnology, Faculty of Life Sciences, Tel-Aviv University, Ramat Aviv 69978, Tel-Aviv, Israel.
Solomon
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B
D002268:Carboxypeptidases / Q000737:chemistry*; D043422:Carboxypeptidases A; D002413:Cations, Divalent; D004795:Enzyme Stability; D005456:Fluorescent Dyes / Q000737:chemistry; D013054:Spectrophotometry, Atomic; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D013816:Thermodynamics; D015032:Zinc / Q000737:chemistry*
D002413:Cations, Divalent; D005456:Fluorescent Dyes; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; D015032:Zinc