Crystallization and initial X-ray diffraction of BtuB, the integral membrane cobalamin transporter of Escherichia coli.
BtuB, the cobalamin transporter from Escherichia coli, has been overexpressed, purified and crystallized. The purified protein was solubilized in n-octyl tetraoxyethylene (C(8)E(4)) and was crystallized using sitting-drop vapor diffusion with PEG 3350 and magnesium acetate as precipitants (pH 6.5). Two crystal forms have been obtained. Crystal type I belongs to space group P3(1)21, with unit-cell parameters a = b = 81.6, c = 210.0 A, alpha = beta = 90, gamma = 120 degrees. Crystal type II belongs to space group P3(1)21, with unit-cell parameters a = b = 81.6, c = 226.0 A, alpha = beta = 90, gamma = 120 degrees. Each crystal form contains a monomer in the asymmetric unit. Diffraction for crystal type I extends to 2.0 A and diffraction for crystal type II extends to 2.7 A. Both crystal forms are suitable for structure determination.
Chimento
David P
DP
Department of Microbiology, University of Virginia, Charlottesville, Virginia 22908, USA.
Mohanty
Arun K
AK
Kadner
Robert J
RJ
Wiener
Michael C
MC
D001425:Bacterial Outer Membrane Proteins; D003001:Cloning, Molecular; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000737:chemistry*; D026901:Membrane Transport Proteins; D018000:Receptors, Peptide / Q000096:biosynthesis / Q000737:chemistry*; D014805:Vitamin B 12 / Q000378:metabolism; D014961:X-Ray Diffraction
D001425:Bacterial Outer Membrane Proteins; C044592:BtuB protein, E coli; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D018000:Receptors, Peptide; D014805:Vitamin B 12