Hermann J, Titani K, Ericsson L H, Wade R D, Neurath H, Walsh K A
Biochemistry. 1978 Dec 26;17(26):5672-9. doi: 10.1021/bi00619a013.
This communication presents the strategy and experimental details to prove the amino acid sequence of two large fragments of rabbit muscle glycogen phosphorylase generated by cleavage with cyanogen bromide. These fragments, CB18 and CB15, represent 241 of the 841 residues in the whole molecule. In addition to applying methods of automated liquid phase Edman degradation, techniques of selective immobilization and solid phase Edman degradation are used. One of the two cyanogen bromide fragments (CB15) contains two of the sites of cleavage with hydroxylamine which have proved to be important in the overall strategy of determining the complete sequence of this molecule. Together with the accompanying reports by Koide, A., et al., and Titani, K., et al. ((1978) Biochemistry 17 (first and third papers, respectively, in a series in this issue)), the present communication completes the proof of the amino acid sequence of phosphorylase and provides the basis for examining the relationship between its structure and function.
本通讯介绍了用于证明经溴化氰裂解产生的兔肌肉糖原磷酸化酶两个大片段氨基酸序列的策略和实验细节。这些片段，即CB18和CB15，占整个分子841个残基中的241个。除了应用自动液相埃德曼降解方法外，还使用了选择性固定技术和固相埃德曼降解技术。两个溴化氰片段之一（CB15）含有两个被羟胺裂解的位点，这些位点在确定该分子完整序列的总体策略中已被证明是重要的。与小出晃等人及谷仁等人的相关报告（（1978年）《生物化学》17卷（分别为本期刊一系列文章中的第一篇和第三篇））一起，本通讯完成了磷酸化酶氨基酸序列的证明，并为研究其结构与功能之间的关系提供了基础。
