RIBOSOME-CATALYZED ESTER FORMATION
RIBOSOME-CATALYZED ESTER FORMATION
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DOI：
10.1021/bi00814a013
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发表时间：
1970-01-01
期刊：
影响因子：
2.9
通讯作者：
RICH, A
中科院分区：
文献类型：
--
作者：
FAHNESTOCK, S;NEUMANN, H;RICH, A
普鲁霉素由环化公司和营养生物化学公司提供。氯霉素是Parke Davis和Co.Gougerotin公司赠送的礼物，武田化学工业有限公司的T.Kanzaki博士慷慨地提供了Gougerotin。以扑霉素为原料，经亚硝酸脱氨反应制得A-羟基嘌呤霉素。将100 mg二盐酸嘌呤霉素溶于20毫升冷(0)1n盐酸中。在此基础上，缓慢搅拌，加入20毫升冷的1nNaN02。将混合物在0度下孵化一夜。然后加入氢氧化钠(4毫升，5毫升)，用三个50毫升等份的乙酸乙酯提取混合物。用25mlH20洗涤结合的乙酸乙酯层，并在常温下减压干燥。该物质溶于甲醇，在2 mm厚的硅胶G上，以四氯化碳-甲醇(10：1)为荧光指示剂进行层析。紫外吸收带被定位，并用丙酮洗脱。获得了三条带，其中没有一条是
Materials and MethodsPuromycin was obtained from CycloChemical Co. and Nutritional Biochemicals. Chloramphenicol was a gift from Parke Davis and Co. Gougerotin was generously supplied as a gift by Dr. T. Kanzaki of Takeda Chemical Industries, Ltd.Puromycin Analogs. a-Hydroxypuromycin was obtained by nitrous acid deamination of puromycin. Puromycin dihydrochloride (100 mg) was dissolved in 20 ml of cold (0) 1 n HC1. To this was added, slowlywith stirring, 20 ml of cold 1 n NaN02. The mixture was incubated overnight at 0. NaOH (4 ml, 5 n) was then added, and the mixture extracted with three 50-ml aliquots of ethyl acetate. Combined ethyl acetate layers were washed with 25 ml of H20 and dried at room temperature under reduced pressure. The material was dissolved in methanol and subjected to chromatography on a 2-mm thick layer of silica gel G with fluores-cent indicator (Analtech) in carbon tetrachloride-methanol (10: 1). Ultraviolet-absorbing bands were located and eluted with acetone. Three bands were obtained, none of which