中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
Expression, purification, and characterization of a new heterotetramer structure of leucyl-tRNA synthetase from Aquifex aeolicus in Escherichia coli

文献类型:期刊论文

作者Olieric, N; Bey, G; Nierengarten, H; Wang, ED; Moras, D; Eriani, G; Cavarelli, J
刊名PROTEIN EXPRESSION AND PURIFICATION
出版日期2006
卷号47期号:1页码:42013
关键词leucyl-tRNA synthetase quaternary structure
通讯作者Cavarelli, J (reprint author), Univ Louis Pasteur Strasbourg 1, CNRS INSERM, Inst Genet & Biol Mol & Cellulaire, Dept Biol & Genom Struct,UMR 7104, 1 Rue Laurent Fries, F-67404 Illkirch Graffenstaden, France.,cava@igbmc.u-strasbg.fr
英文摘要Aminoacyl-tRNA synthetases are key players in the interpretation of the genetic code. They constitute a textbook example of multidomain proteins including insertion and terminal functional modules appended to one of the two class-specific active site domains. The non-catalytic domains usually have distinct roles in the aminoacylation reaction. Aquifex aeolicus leucyl-tRNA synthetase (LeuRS) is composed of a separated catalytic site and tRNA anticodon-binding site, which would represent one of the closest relics of the primordial aminoacyl-tRNA synthetase. Moreover, the essential catalytic site residues are split into the two different subunits. In all other class-I aminoacyl-tRNA synthetases, those two functional polypeptides are nowadays fused into a single protein chain. In this work, we report the isolation and the characterization, in Escherichia coli, of a novel oligomeric form (alpha beta)(2) for A. aeolicus LeuRS, which is present in addition to the alpha beta heterodimer. A. aeolicus (alpha beta)(2) LeuRS has been characterized by biochemical and biophysical methods. Native gel electrophoresis, mass spectrometry, analytical ultracentrifugation, and kinetic analysis confirmed that the (alpha beta)(2) enzyme was a stable and active entity. By mass spectrometry we confirmed that the heterodimer up can bind one tRNA(Leu) molecule whereas the heterotetramer (alpha beta)(2) can bind two tRNA(Leu) molecules. Active site titration and aminoacylation assays showed that two functional active sites are found per heterotetramer, suggesting that this molecular species might exist and be active in vivo. All those data suggest that the existence of the heterotetramer is certainly not an artifact of overexpression in E. coli. (c) 2005 Elsevier Inc. All rights reserved.
学科主题Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology
类目[WOS]Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]MASS-SPECTROMETRY ; SACCHAROMYCES-CEREVISIAE ; GENETIC-CODE ; QUATERNARY STRUCTURE ; COMPLEXES ; EVOLUTION ; SELECTION ; SEQUENCE ; BINDING ; ACIDS
收录类别SCI
语种英语
WOS记录号WOS:000237604200001
版本出版稿
源URL[http://202.127.25.143/handle/331003/1646]  
专题上海生化细胞研究所_上海生科院生化细胞研究所
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Olieric, N,Bey, G,Nierengarten, H,et al. Expression, purification, and characterization of a new heterotetramer structure of leucyl-tRNA synthetase from Aquifex aeolicus in Escherichia coli[J]. PROTEIN EXPRESSION AND PURIFICATION,2006,47(1):42013.
APA Olieric, N.,Bey, G.,Nierengarten, H.,Wang, ED.,Moras, D.,...&Cavarelli, J.(2006).Expression, purification, and characterization of a new heterotetramer structure of leucyl-tRNA synthetase from Aquifex aeolicus in Escherichia coli.PROTEIN EXPRESSION AND PURIFICATION,47(1),42013.
MLA Olieric, N,et al."Expression, purification, and characterization of a new heterotetramer structure of leucyl-tRNA synthetase from Aquifex aeolicus in Escherichia coli".PROTEIN EXPRESSION AND PURIFICATION 47.1(2006):42013.

入库方式: OAI收割

来源:上海生物化学与细胞生物学研究所

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