Study of the thermal denaturation of ribonuclease A by differential thermal analysis and susceptibility to proteolysis.
1. The thermally induced change in conformation of ribonuclease A in solution was investigated by differential thermal analysis and the susceptibility of the enzyme to proteolytic digestion by ficin. 2. A transition with a mid-point of 60.5 degrees C at pH4.2 was observed directly by differential thermal analysis and shown to be a property of the native structure. 3. At pH4.2 ribonuclease A is susceptible to ficin digestion at 60 degrees C but not at 18 degrees C. 4. Chromatographic analysis of the digestion products reveals that transient active intermediates are produced during the digestion. 5. Three of these intermediates were purified and partially characterized. 6. The nature of those sections of the ribonuclease molecule that are involved in the thermal transition is discussed.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D004789:Enzyme Activation; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011489:Protein Denaturation; D012260:Ribonucleases / Q000032:analysis; D013696:Temperature
D000596:Amino Acids; D012260:Ribonucleases