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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Interstice mutations that block site-to-site translocation of a misactivated amino acid bound to a class I tRNA synthetase.
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Interstice mutations that block site-to-site translocation of a misactivated amino acid bound to a class I tRNA synthetase.

机译：间隙突变可阻止与I类tRNA合成酶结合的失活氨基酸的位点转移。

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摘要

Class I aminoacyl-tRNA synthetases catalyze editing reactions that prevent ambiguity from entering the genetic code. Misactivated amino acids are translocated in cis from the active site for aminoacylation to the center for editing, located approximately 30 A away. Mutational analysis has functionally separated the two sites by creating mutations that disrupt the catalytic center for editing but not for aminoacylation and vice versa. What is not known is whether translocation per se can be disrupted without an effect on either catalytic center. Here we describe mutations in a presumptive "hinge region" of isoleucyl-tRNA synthetase that is situated between the two sites. Interstice mutations had little or no effect on either catalytic center. In contrast, the same specific mutations disrupted translocation. Thus, with these mutations all three functions, translocation, catalysis of aminoacylation, and editing, have been mutationally separated. The results are consistent with translocation involving a hinge-region conformational shift that does not perturb the two catalytic centers.
机译：I类氨酰基-tRNA合成酶催化编辑反应，从而防止歧义进入遗传密码。失活的氨基酸以顺式方式从活性位点进行氨基酰化转移至位于约30 A处的编辑中心。突变分析通过创建突变来破坏两个催化中心的功能，从而破坏催化中心进行编辑，而不破坏氨基酰化作用，反之亦然。还不知道易位本身是否可以在不影响任何催化中心的情况下被破坏。在这里，我们描述了位于两个位点之间的异亮氨酰-tRNA合成酶的推定“铰链区”中的突变。间隙突变对两个催化中心几乎没有影响。相反，相同的特定突变破坏了易位。因此，利用这些突变，所有三个功能，易位，氨基酰化的催化和编辑已被突变分离。结果与涉及不干扰两个催化中心的铰链区构象移位的易位一致。

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