Wallace C J, Harris D E
Biochem J. 1984 Feb 1;217(3):589-94. doi: 10.1042/bj2170589.
We have re-examined the acetimidylation and subsequent deprotection of cytochrome c by published methods in the light of recent findings on the tendency of protein acetimidylation reactions to yield side products of differing net charges. We find that the protection methods do indeed yield a mixture of products, some of which have considerably diminished biological activity. Our observations support a postulated mechanism for the generation of side products, and we have been able to identify the major factor responsible for their formation by published methods. The deprotection method appears to be free of side reactions. We describe a new procedure for acetimidylation that will produce fully N-epsilon-acetimidylated cytochrome c. This derivative, lacking detectable side products and having good biological activity, is useful for structure-function studies and as an intermediate in the semisynthesis of cytochrome c analogues.
鉴于近期有关蛋白质乙酰亚胺化反应产生不同净电荷副产物倾向的研究结果，我们重新审视了已发表方法中细胞色素c的乙酰亚胺化及其后续脱保护过程。我们发现，保护方法确实会产生产物混合物，其中一些产物的生物活性显著降低。我们的观察结果支持了一种关于副产物生成的假设机制，并且我们已经能够确定通过已发表方法导致其形成的主要因素。脱保护方法似乎没有副反应。我们描述了一种新的乙酰亚胺化程序，该程序将产生完全N-ε-乙酰亚胺化的细胞色素c。这种衍生物没有可检测到的副产物且具有良好的生物活性，可用于结构-功能研究以及作为细胞色素c类似物半合成的中间体。
