Total synthesis of bovine pancreatic ribonuclease A. Part 6. Synthesis of RNase A with full enzymic activity
Total synthesis of bovine pancreatic ribonuclease A. Part 6. Synthesis of RNase A with full enzymic activity
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DOI：
10.1039/p19810000831
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发表时间：
1981
期刊：
影响因子：
--
通讯作者：
N. Fujii;H. Yajima
中科院分区：
文献类型：
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作者：
N. Fujii;H. Yajima
以Z(OMe)-(RNase 21-124)- obzl、Z(OMe)-(RNase 9-20)- nhnh2和Z-(RNase 1-8)- nhnh2三个片段为缩合物，合成了保护性牛胰腺RNase A。用甲烷磺酸去除保护基团，通过空气氧化建立二硫桥，并在cm -纤维素上通过亲和层析和离子交换层析纯化，得到了与天然RNase a在化学和酶学标准上无法区分的合成蛋白。因此，首次完成了具有充分酶活性的蛋白质的明确合成。
Protected bovine pancreatic RNase A was synthesized by azide condensation of the three fragments Z(OMe)-(RNase 21–124)-OBzl, Z(OMe)-(RNase 9–20)-NHNH2, and Z-(RNase 1–8)-NHNH2. After removal of the protecting groups by methanesulphonic acid, establishment of the disulphide bridges by aerial oxidation, and purification by affinity chromatography and ion-exchange chromatography on CM-cellulose, a synthetic protein was obtained which was indistinguishable by chemical and enzymatic criteria from natural RNase A. Comparable results were also obtained when HF was used as a deprotecting reagent. The unambiguous synthesis of a protein having full enzymic activity has thus been accomplished for the first time.