Overexpression, purification, crystallization and preliminary X-ray studies of Vibrio cholerae EpsG.
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.
Jens
Jason
J
Department of Microbiology and Molecular Genetics and Center for Microbial Pathogenesis, Michigan State University, East Lansing, MI 48824, USA.
Raghunathan
Kannan
K
Vago
Frank
F
Arvidson
Dennis
D
D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification*; D003460:Crystallization; D003625:Data Collection; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D006639:Histidine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D010957:Plasmids; D011994:Recombinant Proteins / Q000302:isolation & purification; D013223:Statistics as Topic; D013696:Temperature; D014734:Vibrio cholerae / Q000235:genetics* / Q000302:isolation & purification*; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D006639:Histidine; D008715:Methionine