Open AccessPurification, crystallization and initial X‐ray crystallographic analysis of the putative GTPase PH0525 from Pyrococcus horikoshii OT3
Author(s) -
Lokanath Neratur K.,
Yamamoto Hitoshi,
Matsunaga Emiko,
Sugahara Mitsuaki,
Kunishima Naoki
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105027752
Subject(s) - pyrococcus horikoshii , gtpase , crystallization , monomer , crystallography , pyrococcus furiosus , escherichia coli , biology , chemistry , biochemistry , crystal structure , archaea , gene , polymer , organic chemistry
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 Å have been collected and processed in space groups P 2 1 and C 222 1 , respectively. Assuming the presence of one monomer per asymmetric unit gives V M values of 2.6 and 2.4 Å 3 Da −1 for the P 2 1 and C 222 1 forms, respectively, which is consistent with dynamic light‐scattering experiments, which show a monomeric state of the protein in solution.