发夹状核酶中的核碱基催化作用。
Nucleobase catalysis in the hairpin ribozyme.
作者信息
Wilson Timothy J, Ouellet Jonathan, Zhao Zheng-Yun, Harusawa Shinya, Araki Lisa, Kurihara Takushi, Lilley David M J
机构信息
Cancer Research UK Nucleic Acid Structure Research Group, The University of Dundee.
出版信息
RNA. 2006 Jun;12(6):980-7. doi: 10.1261/rna.11706. Epub 2006 Apr 6.
Abstract
RNA catalysis is important in the processing and translation of RNA molecules, yet the mechanisms of catalysis are still unclear in most cases. We have studied the role of nucleobase catalysis in the hairpin ribozyme, where the scissile phosphate is juxtaposed between guanine and adenine bases. We show that a modified ribozyme in which guanine 8 has been substituted by an imidazole base is active in both cleavage and ligation, with ligation rates 10-fold faster than cleavage. The rates of both reactions exhibit bell-shaped dependence on pH, with pK(a) values of 5.7 +/- 0.1 and 7.7 +/- 0.1 for cleavage and 6.1 +/- 0.3 and 6.9 +/- 0.3 for ligation. The data provide good evidence for general acid-base catalysis by the nucleobases.
摘要
RNA催化在RNA分子的加工和翻译过程中很重要，但在大多数情况下，催化机制仍不清楚。我们研究了核碱基催化在发夹状核酶中的作用，其中可裂解的磷酸基团位于鸟嘌呤和腺嘌呤碱基之间。我们发现，将鸟嘌呤8用咪唑碱基取代的修饰核酶在切割和连接反应中均具有活性，连接速率比切割速率快10倍。两种反应的速率对pH均呈钟形依赖关系，切割反应的pK(a)值为5.7±0.1和7.7±0.1，连接反应的pK(a)值为6.1±0.3和6.9±0.3。这些数据为核碱基的一般酸碱催化提供了有力证据。
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