Initiating a structural study of 2-keto-3-deoxy-6-phosphogluconate aldolase from Escherichia coli
Buchanan LV., Mehta N., Pocivavsek L., Niranjanakumari S., Toone EJ., Naismith JH.
2-Keto-3-deoxy-6-phosphogluconate aldolase (KDPG aldolase, E.C. 126.96.36.199) 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 Å on station PX9.6 at the Daresbury synchrotron. The crystal belongs to space group P212121, with unit-cell parameters a = 53.2, b = 77.9, c = 146.8 Å.