Two essential regions for tRNA recognition in Bacillus subtilis tryptophanyl-tRNA synthetase
文献类型：期刊论文
| 作者 | Jia, J; Xu, F; Chen, XL; Chen, L; Jin, YX; Wang, DTP |
| 刊名 | BIOCHEMICAL JOURNAL
 |
| 出版日期 | 2002 |
| 卷号 | 365期号:1页码:749-756 |
| 关键词 | acceptor minihelix anticodon microhelix tRNA(Trp)-binding domain |
| 通讯作者 | Jin, YX (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, State Key Lab Mol Biol, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China., |
| 英文摘要 | Bacillus subtilis tryptophanyl-tRNA synthetase (TrpRS) is a homodimeric enzyme. A model for its ability to recognize tRNA(Trp) in B. subtilis was proposed by using computer modelling. This was based on the the fact that there is high homology among bacterial TrpRSs [Chen, Jiang, Jin and Wang (200 1) Acta Biochim. Biophys. Sinica 33, 687-690], in which the enzyme dimer binds to two tRNA(Trp) molecules and each tRNA(Trp) is bound to two different domains across the surface of the dimer. In this work, three deletion mutants of TrpRS were constructed and their products were purified. After determining the kinetic parameters of the mutants in the two-step reaction, it was found that the relative activities of wild-type and mutant enzymes had changed little in the ATP-pyrophosphate exchange reaction. In contrast, the activities of three mutant proteins were much decreased in the tRNA(Trp) aminoacylation assay. Deletion of residues 108-122 and residues 234-238 caused 44% and 80% reductions in the activity, respectively. When both regions were deleted, the aminoacylation activity of the TrpRS mutant was too low to be determined using tRNA(Trp) at the limiting concentration. Gel-retardation assays showed that the acceptor minihelix and the anticodon microhelix were recognized by the domains of TrpRS spanning residues 108-122 and residues 234-238 respectively. In addition, the deletion of amino acids 234-238 affected the normal induced expression of TrpRS at 37 degreesC. In conclusion, residues 108-122 and 234-238 were found essential for tRNA(Trp) recognition. |
| 学科主题 | Biochemistry & Molecular Biology |
| 类目[WOS] | Biochemistry & Molecular Biology |
| 关键词[WOS] | TRANSFER-RNA-SYNTHETASE ; PHENYLALANINE TRANSFER-RNA ; HIGH-LEVEL EXPRESSION ; ESCHERICHIA-COLI ; 3-DIMENSIONAL STRUCTURE ; GENETIC-CODE ; AMINOACYLATION ; SEQUENCE ; DOMAIN ; HOMOLOGY |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000177365400019 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2568]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Jia, J,Xu, F,Chen, XL,et al. Two essential regions for tRNA recognition in Bacillus subtilis tryptophanyl-tRNA synthetase[J]. BIOCHEMICAL JOURNAL,2002,365(1):749-756. |
| APA | Jia, J,Xu, F,Chen, XL,Chen, L,Jin, YX,&Wang, DTP.(2002).Two essential regions for tRNA recognition in Bacillus subtilis tryptophanyl-tRNA synthetase.BIOCHEMICAL JOURNAL,365(1),749-756. |
| MLA | Jia, J,et al."Two essential regions for tRNA recognition in Bacillus subtilis tryptophanyl-tRNA synthetase".BIOCHEMICAL JOURNAL 365.1(2002):749-756. |
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