Efficient segmental isotope labeling of multi-domain proteins using Sortase A
文献类型：期刊论文
| 作者 | Freiburger, Lee1,2,3; Sonntag, Miriam1,2,3; Hennig, Janosch1,2,3; Li, Jian4; Zou, Peijian1,2,3,4; Sattler, Michael1,2,3,4 |
| 刊名 | JOURNAL OF BIOMOLECULAR NMR
 |
| 出版日期 | 2015-09-01 |
| 卷号 | 63期号:1页码:1-8 |
| 关键词 | Protein ligation Sortase A Multi-domain proteins Segmental isotope labeling Protein expression |
| 英文摘要 | NMR studies of multi-domain protein complexes provide unique insight into their molecular interactions and dynamics in solution. For large proteins domain-selective isotope labeling is desired to reduce signal overlap, but available methods require extensive optimization and often give poor ligation yields. We present an optimized strategy for segmental labeling of multi-domain proteins using the S. aureus transpeptidase Sortase A. Critical improvements compared to existing protocols are (1) the efficient removal of cleaved peptide fragments by centrifugal filtration and (2) a strategic design of cleavable and non-cleavable affinity tags for purification. Our approach enables routine production of milligram amounts of purified segmentally labeled protein for NMR and other biophysical studies. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine ; Technology |
| 类目[WOS] | Biochemistry & Molecular Biology ; Spectroscopy |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Spectroscopy |
| 关键词[WOS] | STAPHYLOCOCCUS-AUREUS SORTASE ; MOLECULAR-WEIGHT PROTEINS ; SOLUTION NMR ; SUPRAMOLECULAR STRUCTURES ; STRUCTURAL-ANALYSIS ; SURFACE-PROTEINS ; CELL-WALL ; IN-VITRO ; RNA ; LIGATION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000361612700001 |
| 源URL | [http://124.16.173.210/handle/834782/1574]  |
| 专题 | 天津工业生物技术研究所_蛋白质工程实验室 邹培建_期刊论文 |
| 作者单位 | 1.Helmholtz Zentrum Munchen, Inst Biol Struct, D-85764 Neuherberg, Germany 2.Tech Univ Munich, CIPSM, D-85747 Garching, Germany 3.Tech Univ Munich, Dept Chem, D-85747 Garching, Germany 4.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China |
| 推荐引用方式 GB/T 7714 | Freiburger, Lee,Sonntag, Miriam,Hennig, Janosch,et al. Efficient segmental isotope labeling of multi-domain proteins using Sortase A[J]. JOURNAL OF BIOMOLECULAR NMR,2015,63(1):1-8. |
| APA | Freiburger, Lee,Sonntag, Miriam,Hennig, Janosch,Li, Jian,Zou, Peijian,&Sattler, Michael.(2015).Efficient segmental isotope labeling of multi-domain proteins using Sortase A.JOURNAL OF BIOMOLECULAR NMR,63(1),1-8. |
| MLA | Freiburger, Lee,et al."Efficient segmental isotope labeling of multi-domain proteins using Sortase A".JOURNAL OF BIOMOLECULAR NMR 63.1(2015):1-8. |
入库方式： OAI收割
来源：天津工业生物技术研究所
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