Purification, crystallization and initial X-ray crystallographic analysis of the putative GTPase PH0525 from Pyrococcus horikoshii OT3

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Neratur K LokanathNaoki Kunishima

Abstract

GTPases are involved in diverse cellular functions including cell proliferation, cytoskeleton organization and intracellular traffic. The putative GTPase PH0525 from Pyrococcus horikoshii OT3 has been overexpressed in Escherichia coli and purified. Two distinct crystal forms were grown by the microbatch method at 291 K using a very high protein concentration (80 mg ml(-1)). Native data sets extending to resolutions of 2.3 and 2.4 A have been collected and processed in space groups P2(1) and C222(1), respectively. Assuming the presence of one monomer per asymmetric unit gives VM values of 2.6 and 2.4 A3 Da(-1) for the P2(1) and C222(1) forms, respectively, which is consistent with dynamic light-scattering experiments, which show a monomeric state of the protein in solution.

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