An insertion mutant of LeuRS with 116 amino acid residues has full activity
文献类型：期刊论文
| 作者 | Huang, Y; Ling, C; Li, T; Tong, GL; Wang, ED |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2003 |
| 卷号 | 35期号:3页码:225-229 |
| 关键词 | E. coli leucyl-tRNA synthetase insertion mutant expression and purification activity |
| 通讯作者 | , |
| 英文摘要 | Escherichia colt leucyl-tRNA synthetase (LeuRS) belongs to class I aminoacyl-tRNA synthetases. It consists of 860 amino acid residues and catalyzes the leucylation of tRNA(leu). An insertion of its 253-368 peptide fragment between 368 to 369 in CP1 domain of this enzyme was shown to maintain the activity of the enzyme, and the insertion mutant was named as LeuRS-C. Because the insertion mutant of LeuRS was sensitive to operation of the purification, a plasmid containing the gene encoding LeuRS with His(6)-tag at its N-terminus was constructed to facilitate the purification of His(6)-LeuRS-C through one-step affinity chromatography on Ni-NTA column. The purified His(6)-LeuRS-C had full activity as the native LeuRS with His-tag at the N-terminus (His(6)-LeuRS), although the mutant enzyme had an insertion of 116 amino acid residues. The kinetic parameters of His(6)-LeuRS-C were determined. The secondary structure estimated by CD spectrum and thermal stability of the insertion mutant was compared with those of His(6)-LeuRS, respectively. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | TRANSFER-RNA-SYNTHETASE ; ESCHERICHIA-COLI ; TRNA(LEU) ISOACCEPTORS ; DISCRIMINATION ; PURIFICATION ; SEQUENCE ; PROTEIN |
| 收录类别 | SCI |
| 语种 | 中文 |
| WOS记录号 | WOS:000181873400003 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2268]  |
| 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 | Huang, Y,Ling, C,Li, T,et al. An insertion mutant of LeuRS with 116 amino acid residues has full activity[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2003,35(3):225-229. |
| APA | Huang, Y,Ling, C,Li, T,Tong, GL,&Wang, ED.(2003).An insertion mutant of LeuRS with 116 amino acid residues has full activity.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,35(3),225-229. |
| MLA | Huang, Y,et al."An insertion mutant of LeuRS with 116 amino acid residues has full activity".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 35.3(2003):225-229. |
入库方式： OAI收割
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