Crystallization and preliminary X-ray analysis of glucose dehydrogenase from Haloferax mediterranei.
Glucose dehydrogenase (E.C. 1.1.1.47; GlcDH) from Haloferax mediterranei has been overexpressed in Escherichia coli, solubilized by the addition of 8 M urea and refolded by rapid dilution. The protein has been purified by conventional techniques and crystallized by the hanging-drop vapour-diffusion method using sodium citrate as the precipitant. Two crystal forms representing the free enzyme and the binary complex with NADP(+) grow under these conditions. Crystals of form I diffract to beyond 3.5 A resolution and belong to the hexagonal space group P622, with unit-cell parameters a = b = 89.1, c = 214.6 A, alpha = beta = 90, gamma = 120 degrees. Crystals of form II diffract to greater than 2.0 A and belong to the orthorhombic space group I222 or I2(1)2(1)2(1), with unit-cell parameters a = 61.8, b = 110.9, c = 151.7 A, alpha = beta = gamma = 90 degrees. Calculated values for V(M) and consideration of the packing for both crystal forms suggests that the asymmetric units in both crystal forms contain a monomer.
Ferrer
J
J
Departamento de Agroquímica y Bioquímica, Facultad de Ciencias, Universidad de Alicante, Ap. 99, E-03080 Alicante, Spain.
Fisher
M
M
Burke
J
J
Sedelnikova
S E
SE
Baker
P J
PJ
Gilmour
D J
DJ
Bonete
M J
MJ
Pire
C
C
Esclapez
J
J
Rice
D W
DW
D003460:Crystallization; D018360:Crystallography, X-Ray; D042843:Glucose 1-Dehydrogenase; D005948:Glucose Dehydrogenases / Q000737:chemistry* / Q000235:genetics; D019623:Haloferax mediterranei / Q000201:enzymology*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry
D011994:Recombinant Proteins; D005948:Glucose Dehydrogenases; D042843:Glucose 1-Dehydrogenase