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首页> 外文期刊>Biochemistry >From ribonuclease A toward bovine seminal ribonuclease: a step by step thermodynamic analysis
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From ribonuclease A toward bovine seminal ribonuclease: a step by step thermodynamic analysis

机译：从核糖核酸酶A到牛精子核糖核酸酶：逐步热力学分析

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摘要

A proline, a leucine, and two cysteine residues, introduced at positions 19, 28, 31, and 32 of bovine pancreatic RNase A, i.e. the positions occupied by these residues in the subunit of bovine seminal RNase, the only dimeric RNase of the pancreatic-type superfamily, transform monomeric RNase A into a dimeric RNase, endowed with the same ability of BS-RNase of swapping its N-terminal segments. The thermodynamic consequences of the progressive introduction of these four residues into RNase A polypeptide chain have been studied by comparing the temperature- and urea-induced denaturation of three mutants of RNase A with that of a stable monomeric derivative of BS-RNase. The denaturation processes proved reversible for all proteins, and well represented by the two-state N<-->D transition model. The progressive introduction of the four residues into RNase A led to a gradual shift of the protein stability toward that characteristic of monomeric BS-RNase, which, in turn, is markedly less stable than RNase A with respect to both temperature- and urea-induced denaturation. On the other hand, the thermal stability of a dimeric active mutant of RNase A is found to approach that of wild-type seminal RNase.
机译：在牛胰腺RNase A的19、28、31和32位引入的脯氨酸，亮氨酸和两个半胱氨酸残基，即这些残基在牛精浆RNase的亚基中占据的位置，这是胰腺唯一的二聚RNase -型超家族，将单体RNase A转化为二聚RNase，赋予BS-RNase交换其N末端片段的相同能力。通过比较温度和尿素诱导的RNase A的三个突变体与稳定的BS-RNase单体衍生物的变性，研究了将这四个残基逐步引入RNase A多肽链的热力学后果。证明变性过程对于所有蛋白质都是可逆的，并且由两态N--D过渡模型很好地表示。四个残基的逐步引入到RNase A中，导致蛋白质的稳定性逐渐向单体BS-RNase的特征转变，这反过来，在温度和尿素诱导下，其稳定性均明显低于RNaseA。变性。另一方面，发现RNase A的二聚活性突变体的热稳定性接近野生型精浆RNase的热稳定性。

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