PARTIAL DIGESTION OF TRANSFER RNA-AMINOACYL-TRANSFER RNA-SYNTHETASE COMPLEXES WITH COBRA VENOM RIBONUCLEASE
PARTIAL DIGESTION OF TRANSFER RNA-AMINOACYL-TRANSFER RNA-SYNTHETASE COMPLEXES WITH COBRA VENOM RIBONUCLEASE
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DOI：
10.1021/bi00507a055
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发表时间：
1981-01-01
期刊：
影响因子：
2.9
通讯作者：
EBEL, JP
中科院分区：
文献类型：
--
作者：
FAVOROVA, OO;FASIOLO, F;EBEL, JP
转移RNA [tRNA]分子在5“或3”末端用32 P标记，并用眼镜蛇毒RNase消化，切割双链区域。通过高分辨率凝胶电泳分析酵母tRNAPhe和tRNAVal的产物。在游离状态下，tRNA主要在受体和反密码子茎中被切割。T.PSI.有轻微割伤。tRNAVal中的茎。通过确定被蛋白质屏蔽的tRNA区域，研究了与其同源合成酶相互作用的区域的地形。在tRNAVal的反密码子和受体茎中发现近100%的保护。在tRNAPhe中，只有反密码子的茎被保护。研究了tRNAPhe和牛胰腺中的N-乙酰基-tRNA合成酶之间的非同源相互作用。牛肉酶没有保护tRNAPhe，尽管事实上，有效的misaminoacylation发生。对每个tRNA合成酶复合物获得的屏蔽图案与这些复合物的直接UV交联实验的结果进行了比较。
Transfer RNA [tRNA] molecules were labeled with 32P at the 5'' or 3'' end and digested with cobra venom RNase, which cut double-stranded regions. The products of yeast tRNAPhe and tRNAVal were analyzed by high-resolution gel electrophoresis. In the free state, tRNA were cut predominantly in the acceptor and anticodon stems. Minor cuts occurred in the T.PSI. stem in tRNAVal. The topography of zones interacting with their cognate synthetases was studied by determining tRNA regions shielded by protein. Nearly 100% protection was found in anticodon and acceptor stem of tRNAVal. In tRNAPhe, only the stem of the anticodon was protected. Noncognate interactions between tRNAPhe and tryptophanyl-tRNA synthetase from beef pancreas were examined. Beef enzyme did not protect tRNAPhe despite the fact that efficient misaminoacylation occurred. The pattern of shielding obtained for each tRNA-synthetase complex was compared with results of direct UV cross-linking experiments with these complexes.