Jens Jason, Raghunathan Kannan, Vago Frank, Arvidson Dennis
Department of Microbiology and Molecular Genetics and Center for Microbial Pathogenesis, Michigan State University, East Lansing, MI 48824, USA.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Jun 1;65(Pt 6):629-31. doi: 10.1107/S1744309109017163. Epub 2009 May 23.
EpsG is the major pseudopilin protein of the Vibrio cholerae type II secretion system. An expression plasmid that encodes an N-terminally truncated form of EpsG with a C-terminal noncleavable His tag was constructed. Recombinant EpsG was expressed in Escherichia coli; the truncated protein was purified and crystallized by hanging-drop vapor diffusion against a reservoir containing 6 mM zinc sulfate, 60 mM MES pH 6.5, 15% PEG MME 550. The crystals diffracted X-rays to a resolution of 2.26 A and belonged to space group P2(1), with unit-cell parameters a = 88.61, b = 70.02, c = 131.54 A.
EpsG是霍乱弧菌II型分泌系统的主要假菌毛蛋白。构建了一个表达质粒，该质粒编码带有C端不可切割His标签的N端截短形式的EpsG。重组EpsG在大肠杆菌中表达；截短的蛋白经纯化后，通过悬滴气相扩散法，以含有6 mM硫酸锌、60 mM MES pH 6.5、15%聚乙二醇甲醚550的贮液进行结晶。晶体的X射线衍射分辨率为2.26 Å，属于空间群P2(1)，晶胞参数a = 88.61、b = 70.02、c = 131.54 Å。
