通过差示热分析研究核糖核酸酶A的热变性及其对蛋白水解的敏感性。
Study of the thermal denaturation of ribonuclease A by differential thermal analysis and susceptibility to proteolysis.
作者信息
Winchester B G, Mathias A P, Rabin B R
出版信息
Biochem J. 1970 Apr;117(2):299-307. doi: 10.1042/bj1170299.
Abstract
- 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.
摘要
- 通过差示热分析以及该酶对无花果蛋白酶蛋白水解消化的敏感性，研究了溶液中核糖核酸酶A构象的热诱导变化。2. 通过差示热分析直接观察到在pH4.2时中点温度为60.5摄氏度的转变，并表明这是天然结构的一种特性。3. 在pH4.2时，核糖核酸酶A在60摄氏度下易被无花果蛋白酶消化，但在18摄氏度下不易被消化。4. 对消化产物的色谱分析表明，消化过程中会产生瞬时活性中间体。5. 其中三种中间体被纯化并进行了部分表征。6. 讨论了核糖核酸酶分子中参与热转变的那些区域的性质。
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