鼠疫杆菌耶尔森氏菌分泌的一种关键毒力因子YopM的过量生产、纯化、结晶及初步X射线衍射分析

Overproduction, purification, crystallization and preliminary X-ray diffraction analysis of YopM, an essential virulence factor extruded by the plague bacterium Yersinia pestis.

作者信息

Evdokimov A G, Anderson D E, Routzahn K M, Waugh D S

机构信息

Protein Engineering Section, National Cancer Institute - Frederick Cancer Research and Development Center, PO Box B, Frederick, MD 21702, USA.

出版信息

Acta Crystallogr D Biol Crystallogr. 2000 Dec;56(Pt 12):1676-9. doi: 10.1107/s0907444900013378.

DOI:10.1107/s0907444900013378

PMID:11092944

Abstract

A recombinant form of Yersinia pestis YopM with a C-terminal polyhistidine affinity tag has been overproduced in Escherichia coli, purified to homogeneity and crystallized using the hanging-drop vapor-diffusion technique. Several different crystal forms were obtained. The most suitable crystals for X-ray diffraction belonged to space groups P4(2)22 (unit-cell parameters a = 109.36, b = 109.36, c = 101.50 A) and C222(1) (unit-cell parameters a = 71.73, b = 121. 85, c = 189.79 A). With a synchrotron-radiation source, these crystals diffracted to 2.4 and 1.9 A resolution, respectively.

摘要

一种带有C端多组氨酸亲和标签的重组鼠疫耶尔森菌YopM已在大肠杆菌中过量表达，纯化至同质，并采用悬滴气相扩散技术进行结晶。获得了几种不同的晶体形式。最适合X射线衍射的晶体属于空间群P4(2)22（晶胞参数a = 109.36，b = 109.36，c = 101.50 Å）和C222(1)（晶胞参数a = 71.73，b = 121.85，c = 189.79 Å）。利用同步辐射源，这些晶体分别衍射至2.4 Å和1.9 Å分辨率。