Crystallization and preliminary X-ray crystallographic analysis of type II dehydroquinase from Helicobacter pylori.
The enzyme 3-dehydroquinase catalyzes the interconversion of 3-dehydroquinate and 3-dehydroshikimate. The enzymes are classified into two groups, type I and type II, which have different biochemical and biophysical properties and act with different mechanisms. The type II dehydroquinase of Helicobacter pylori, a dodecameric enzyme, was overexpressed in Escherichia coli. The recombinant protein has been crystallized at 296 K using polyethylene glycol (PEG) 4000 as a precipitant. Native X-ray diffraction data have been collected to 2.5 A resolution using synchrotron radiation. The crystals are cubic and belong to the space group P4(2)32, with unit-cell parameters a = b = c = 98.91 A. The asymmetric unit contains one subunit of recombinant type II dehydroquinase, with a corresponding V(M) of 2.18 A(3) Da(-1) and a solvent content of 43.6%.
Kwak
J E
JE
School of Chemistry and Molecular Engineering, Seoul National University, Seoul 151-742, South Korea.
Lee
J Y
JY
Han
B W
BW
Moon J
J
J
Sohn
S H
SH
Suh
S W
SW
D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D016480:Helicobacter pylori / Q000201:enzymology*; D006836:Hydro-Lyases / Q000737:chemistry* / Q000302:isolation & purification; D011092:Polyethylene Glycols; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D017356:Synchrotrons
D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D006836:Hydro-Lyases; C020000:3-dehydroquinate dehydratase