An examination of the utility of photogenerated reagents by using α-chymotrypsin
An examination of the utility of photogenerated reagents by using α-chymotrypsin
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使用 α-胰凝乳蛋白酶检查光生试剂的效用
DOI：
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发表时间：
1974
期刊：
影响因子：
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通讯作者：
J. Knowles
中科院分区：
文献类型：
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作者：
A. Bridges;J. Knowles
作为光生试剂的标记特性的测试，在已知三级结构的蛋白质的芳香族结合位点中光解芳基叠氮化物。从α-胰凝乳蛋白酶与对叠氮[14 C]肉桂酸对硝基苯酯反应中分离并光解了酰基酶。大约60%的酰基在光解和脱酰后共价结合到蛋白质上，标记的酶是无活性的。共价连接的标签位于胰凝乳蛋白酶的C链中，并且有明确的迹象表明，C链的主要标记的胰蛋白酶片段是构成酶的芳香族结合位点的片段。根据α-胰凝乳蛋白酶的已知化学和结构预测的蛋白质分子的该部分的高度标记为光标记方法的实用性提供了令人满意的证实。
As a test of the labelling characteristics of photogenerated reagents, an aryl azide was photolysed in the aromatic-binding locus of a protein of known tertiary structure. The acyl-enzyme derived from the reaction of α-chymotrypsin with the p-nitrophenyl ester of p-azido[14C]cinnamate was isolated and photolysed. About 60% of the acyl group is covalently bound to the protein after photolysis and deacylation, and labelled enzyme is inactive. The covalently attached label is localized in the C chain of chymotrypsin, and there are firm indications that the major labelled tryptic fragment of the C chain is that which constitutes the aromatic-binding locus of the enzyme. The high degree of labelling of that portion of the protein molecule predicted on the basis of the known chemistry and structure of α-chymotrypsin, provides gratifying confirmation of the utility of the photo-labelling method.