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.
<jats:p>2-Keto-3-deoxy-6-phosphogluconate aldolase (KDPG aldolase, E.C. 184.108.40.206) is a member of the pyruvate/phospho<jats:italic>enol</jats:italic>pyruvate 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 <jats:italic>P</jats:italic>2<jats:sub>1</jats:sub>2<jats:sub>1</jats:sub>2<jats:sub>1</jats:sub>, with unit-cell parameters <jats:italic>a</jats:italic> = 53.2, <jats:italic>b</jats:italic> = 77.9, <jats:italic>c</jats:italic> = 146.8 Å.</jats:p>