Preparation of the selenomethionine derivative of tabtoxin resistance protein |
| |
Authors: | Hongzhen He Yu Shao Chen Yuhang Liu Bingbing Cao Zhenbo Jiang Fan Liu Yiwei Tang Hong Liu Jinyuan Zhao Nanming Rao Zihe |
| |
Institution: | (1)Laboratory of Structural Biology & Protein Science Laboratory of MOE, School of Life Science and Engineering, Tsinghua University, Beijing 100084 ,China |
| |
Abstract: | In order to obtain phase information for the X-ray diffraction of tabtoxin resistance protein (TTR) crystal using the MAD phasing method, a selenomethionine (Se-Met) derivative of TTR was overexpressed in E. coli strain M15, with pQE-30 plasmid, through IPTG induction in M9 medium containing Se-Met. The product was purified to an estimated homogeneity of greater than 95% according to SDS-PAGE, by a Ni-NTA metal affinity followed by a Mono Q anion exchange column chromatography. The successful substitution of Se-Met for methionine (Met) was confirmed by MALDI-TOF and ESI-Quadrupole Mass Spectrometry analysis. The derivative crystal was obtained using similar conditions as those for the native. |
| |
Keywords: | Se-Met TTR derivative expression purification mass spectrometry |
本文献已被 SpringerLink 等数据库收录! |
| 点击此处可从《中国科学通报(英文版)》浏览原始摘要信息 |
| 点击此处可从《中国科学通报(英文版)》下载免费的PDF全文 |