Products from the photolysis of diazoacetyl chymotrypsin.
Products from the photolysis of diazoacetyl chymotrypsin.
复制标题
重氮乙酰糜蛋白酶光解的产物。
DOI：
10.1016/s0021-9258(18)96934-6
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发表时间：
1966
期刊：
影响因子：
--
通讯作者：
F. Westheimer
中科院分区：
文献类型：
--
作者：
J. Shafer;P. Baronowsky;R. Laursen;F. Finn;F. Westheimer
1.以对硝基苯基氯甲酸酯和重氮甲烷为原料，合成了一种新的简便的对硝基苯基重氮乙酸酯。2.对硝基苯基重氮乙酸酯与糜蛋白酶按化学计量比反应生成重氮乙酰胰凝乳蛋白酶。重氮乙酰基酶在波长大于320m微米的光下进行光解，得到(A)乙醇酸和再生的胰凝乳蛋白酶和(B)保留重氮乙酰基团的碳原子的修饰蛋白质的混合物。从光解得到的蛋白质的酸解产生O-羧甲基丝氨酸，通过层析和其他比较与真品和氮杂丝氨酸的光解形成的物质进行鉴定。此外，还得到了少量的1-羧甲基组氨酸和Ofo-羧甲基酪氨酸。前者的来源尚不清楚，但后者可能源于重氮乙酰化酶的二聚体的光解。大多数产物可能来自作为重氮酯光解主要产物的卡宾。
1. A new and convenient synthesis ofp-nitrophenyl diazoacetate has been devised fromp-nitrophenyl chloroformate and diazomethane.2.p-Nitrophenyl diazoacetate reacts stoichiometrically with chymotrypsin to yield diazoacetyl chymotrypsin.3. Photolysis of the diazoacetyl enzyme with light of wave length greater than 320 mµ yields a mixture of (a) glycolic acid and regenerated chymotrypsin and (b) modified protein which retains the carbon atoms of the diazoacetyl group.4. Acid hydrolysis of the protein obtained from photolysis yieldsO-carboxymethylserine, identified by chromatographic and other comparisons with an authentic sample and with material formed on photolysis of azaserine.5. In addition, small amounts of 1-carboxymethylhistidine and ofO-carboxymethyltyrosine were obtained. The origin of the former is uncertain, but the latter may arise from photolysis of a dimer of the diazoacetylated enzyme.6. Most of the products presumably arise from a carbene formed as a primary product of the photolysis of the diazoester.