Preparation of the selenome-thionine derivative of tab-toxin resistance protein  

作　　者：HE Hongzhen SHAO Yu CHEN Yuhang LIU Bingbing CAO Zhenbo JIANG Fan LIU Yiwei TANG Hong LIU Jinyuan ZHAO Nanming RAO Zihe 

机构地区：[1]Laboratory of Structural Biology&Protein Science Laboratory of MOE,School of Life Science and Engineering,Tsinghua University,Beijing 100084,China

出　　处：《Chinese Science Bulletin》2001年第15期1304-1307,共4页

基　　金：This work wassupported by the National Natural Science Foundation of China (Grant Nos. 39870174 and 39970155); the National "863" Project (Grant No. 103130306) and the National "973" Project (Grant Nos. G1999075602, G19990II902 and 1998051105).

摘　　要：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.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 inE. 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.

关 键 词：Se-Met TTR DERIVATIVE expression purification mass spectrometry. 

分 类 号：Q51[生物学—生物化学]