Initiating a structural study of 2-keto-3-deoxy-6-phosphogluconate aldolase from Escherichia coli.
2-Keto-3-deoxy-6-phosphogluconate aldolase (KDPG aldolase, E.C. 4.1. 2.14) is a member of the pyruvate/phosphoenolpyruvate aldolase family. It is also a synthetically useful enzyme, capable of catalyzing the stereoselective aldol addition of pyruvate to a range of unnatural electrophilic substrates. The recombinant protein was purified by a two-step HPLC protocol involving anion-exchange and hydrophobic chromatography. Dynamic light-scattering experiments indicated the protein to be monodisperse. Crystals were obtained using the sitting-drop vapour-diffusion method, with PEG 6K as precipitant. Diffraction data were collected on a frozen crystal to a resolution of 2.26 A on station PX9.6 at the Daresbury synchrotron. The crystal belongs to space group P2(1)2(1)2(1), with unit-cell parameters a = 53.2, b = 77.9, c = 146.8 A.
Buchanan
L V
LV
Centre for Biomedical Sciences, North Haugh, The University, St Andrews, Fife KY16 9ST, Scotland.
Mehta
N
N
Pocivavsek
L
L
Niranjanakumari
S
S
Toone
E J
EJ
Naismith
J H
JH
D000446:Aldehyde-Lyases / Q000737:chemistry*; D000595:Amino Acid Sequence; D003460:Crystallization; D004926:Escherichia coli; D008969:Molecular Sequence Data; D011092:Polyethylene Glycols / Q000494:pharmacology; D011487:Protein Conformation; D016958:Pseudomonas putida; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D012645:Selenomethionine / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D014961:X-Ray Diffraction
D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D012645:Selenomethionine; D000446:Aldehyde-Lyases; C100131:phospho-2-keto-3-deoxy-gluconate aldolase