Immobilized carboxypeptidase A as a probe for studying the thermally induced unfolding of bovine pancreatic ribonuclease.
A method for the preparation of Sephadex-immobilized carboxypeptidase A is presented. This form of the enzyme has the same specific activity as the soluble enzyme at room temperature, but retains its activity at higher temperatures (60-70 degrees). This preparation of immobilized carboxypeptidase A was used, as a proteolytic probe, to investigate the thermally induced unfolding of the C-terminus of ribonuclease A. This technique indicates that the C-terminal residues of ribonuclease A do not unfold until the high-temperature region of the thermal transition (as determined by ultraviolet difference spectrophotometry and optical rotation).
D000818:Animals; D002268:Carboxypeptidases; D002417:Cattle; D006358:Hot Temperature; D007700:Kinetics; D008722:Methods; D010179:Pancreas / Q000201:enzymology*; D011485:Protein Binding; D011489:Protein Denaturation; D012260:Ribonucleases; D012995:Solubility
D012260:Ribonucleases; D002268:Carboxypeptidases