Crystallization and preliminary X-ray diffraction studies of beta-phosphoglucomutase from Lactococcus lactus.
Beta-phosphoglucomutase (beta-PGM), a 28 kDa monomer, catalyzes the reversible conversion of beta-D-glucose-1-phosphate to beta-D-glucose-6-phosphate in maltose metabolism in a variety of organisms. Sequence analysis of beta-PGM indicates that it is a member of the haloacid dehalogenase (HAD) enzyme superfamily, which evolved to cleave C-Cl, C-P and C-OP bonds in a variety of substrates. beta-PGM has been crystallized using the hanging-drop method. Diffraction-quality crystals of the native protein have been obtained from two conditions, both belonging to space group P2(1)2(1)2(1), with unit-cell parameters a = 53.67, b = 92.78, c = 111.60 and a = 53.21, b = 57.01, c = 76.11 A. To solve the phase problem, selenomethionine (SeMet) containing beta-PGM crystals have been grown. The SeMet-containing crystals diffract to high resolution only when grown by microseeding with native crystals. A three-wavelength data set has been collected to 2.3 A on crystals of the SeMet-substituted beta-PGM. The structure solution is currently being attempted by the multiwavelength anomalous diffraction (MAD) phasing method.
Lahiri
Sushmita D
SD
Department of Physiology and Biophysics, Boston University School of Medicine, Boston, Massachusetts 02118-2394, USA.
Zhang
Guofeng
G
Radstrom
Peter
P
Dunaway-Mariano
Debra
D
Allen
Karen N
KN
D003460:Crystallization; D018360:Crystallography, X-Ray; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D013294:Lactococcus lactis / Q000201:enzymology*; D010733:Phosphoglucomutase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation
D010733:Phosphoglucomutase