光亲和标记
核糖体
大肠杆菌
化学
A站点
四环素
结合位点
核糖体RNA
生物化学
凝胶电泳
聚丙烯酰胺凝胶电泳
生物物理学
生物
核糖核酸
抗生素
酶
基因
作者
Tayyaba Hasan,Robert A. Goldman,Barry S. Cooperman
标识
DOI:10.1016/0006-2952(85)90610-0
摘要
[3H]Tetracycline (TC) has been shown to photoincorporate into the Escherichia coli ribosome. However, the utility of this process for characterizing the TC binding site on the ribosome is diminished by competing side reactions which also lead to incorporation of radioactivity. In this work we first conducted a detailed study of the labeling processes occurring when ribosomes are irradiated in the presence of [3H]TC with a common, rather low intensity, lamp. On the basis of the results of this study we next explored the usefulness for photoaffinity labeling of the TC site of both irradiation with a high-intensity laser and radioactive, functional TC derivatives having different photochemical properties than TC itself. Labeling patterns determined by polyacrylamide gel electrophoretic analysis of ribosomal proteins extracted from photoaffinity-labeled 30S subunits provided strong evidence that these two approaches offer distinct advantages for characterizing the TC binding site.
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