Thermal stabilization of carboxypeptidase A as a function of pH and ionic milieu.
In this study we investigated the contribution of pH, phosphate anions and salt concentration to the catalytic and structural thermostability of the carboxypeptidase A (CPA). The concentration of 75-100 mM phosphate as well as neutral pH values were found to be optimal for stabilizing CPA at high temperatures. Although moderate concentrations of sodium chloride had no effect on thermal stability, high concentrations of the salt destabilized the enzyme. The experimental results and theoretical analysis suggested that the main contribution to heat stabilization of CPA is related to intramolecular electrostatic interactions and Arginine and/or Lysine are the putative groups able to bind phosphate and stabilize the enzyme molecule against thermal denaturation.
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Department of Molecular Microbiology & Biotechnology, Faculty of Life Sciences, Tel-Aviv University, Ramat Aviv, Israel.
Solomon
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D000818:Animals; D001665:Binding Sites; D002268:Carboxypeptidases / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D043422:Carboxypeptidases A; D002417:Cattle; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D008956:Models, Chemical; D008961:Models, Structural; D010710:Phosphates / Q000378:metabolism / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D013696:Temperature
D010710:Phosphates; D012965:Sodium Chloride; D002268:Carboxypeptidases; D043422:Carboxypeptidases A