Crystallization and preliminary structure analysis of CobE, an essential protein of cobalamin (vitamin B12) biosynthesis.
CobE, a protein implicated in vitamin B12 biosynthesis, from Pseudomonas aeruginosa has been overexpressed in Escherichia coli, purified and crystallized using hanging-drop vapour diffusion. The crystals belong to the primitive orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 31.86, b = 41.07, c = 87.41 A. The diffraction extends to a resolution of 1.9 A. There is one molecule per asymmetric unit and the estimated solvent content is 35%. SeMet-labelled CobE has been prepared and crystallizes under the same conditions as the native protein with diffraction to 1.7 A. The anomalous measurements will be used for phasing.
Vévodová
Jitka
J
Department of Chemistry, University of York, Heslington, York YO10 5YW, England.
Graham
Ross M
RM
Raux
Evelyne
E
Warren
Martin J
MJ
Wilson
Keith S
KS
D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D012645:Selenomethionine; D014805:Vitamin B 12 / Q000096:biosynthesis; D014835:Volatilization; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D012645:Selenomethionine; D014805:Vitamin B 12