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A730环是VS核酶活性位点的一个重要组成部分。

The A730 loop is an important component of the active site of the VS ribozyme.

作者信息

Lafontaine D A, Wilson T J, Norman D G, Lilley D M

机构信息

CRC Nucleic Acid Structure Research Group, Department of Biochemistry, MSI/WTB Complex, The University of Dundee, Dundee DD1 5EH, UK.

出版信息

J Mol Biol. 2001 Sep 28;312(4):663-74. doi: 10.1006/jmbi.2001.4996.

Abstract

The core of the VS ribozyme comprises five helices, that act either in cis or in trans on a stem-loop substrate to catalyse site-specific cleavage. The structure of the 2-3-6 helical junction indicates that a cleft is formed between helices II and VI that is likely to serve as a receptor for the substrate. Detailed analysis of sequence variants suggests that the base bulges of helices II and VI play an architectural role. By contrast, the identity of the nucleotides in the A730 loop is very important for ribozyme activity. The base of A756 is particularly vital, and substitution by any other nucleotide or ablation of the base leads to a major reduction in cleavage rate. However, variants of A756 bind substrate efficiently, and are not defective in global folding. These results suggest that the A730 loop is an important component of the active site of the ribozyme, and that A756 could play a key role in catalysis.

摘要

VS核酶的核心由五个螺旋组成,这些螺旋可对茎环底物进行顺式或反式作用,以催化位点特异性切割。2-3-6螺旋连接的结构表明,在螺旋II和VI之间形成了一个裂缝,该裂缝可能作为底物的受体。对序列变体的详细分析表明,螺旋II和VI的碱基凸起起到了结构作用。相比之下,A730环中核苷酸的身份对核酶活性非常重要。A756的碱基尤为关键,被任何其他核苷酸取代或碱基缺失都会导致切割速率大幅降低。然而,A756的变体能够有效地结合底物,并且在整体折叠方面没有缺陷。这些结果表明,A730环是核酶活性位点的重要组成部分,并且A756可能在催化中发挥关键作用。

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