Open AccessPurification, crystallization and preliminary X‐ray crystallographic analysis of the archaeal phosphoglycerate mutase PH0037 from Pyrococcus horikoshii OT3
Author(s) -
Lokanath Neratur K.,
Kunishima Naoki
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106026121
Subject(s) - pyrococcus horikoshii , phosphoglycerate mutase , crystallography , phosphoglycerate kinase , dimer , crystallization , chemistry , crystal structure , biochemistry , enzyme , glycolysis , organic chemistry
Phosphoglycerate mutases catalyze the interconversion of 2‐phosphoglycerate and 3‐phosphoglycerate in glycolysis and gluconeogenesis pathways. The archaeal phosphoglycerate mutase PH0037 from Pyrococcus horikoshii OT3 has been overexpressed in Escherichia coli and purified. Crystals were obtained using the oil‐microbatch method at 291 K. A native data set extending to a resolution of 2.2 Å has been collected and processed in space group R 32. Assuming the presence of a dimer in the asymmetric unit, the V M value is calculated to be 3.0 Å 3 Da −1 , consistent with the dynamic light‐scattering experiment result, which shows a dimeric state of the protein in solution. Molecular‐replacement trials using the crystal structure of Bacilllus stearothermophilus phosphoglycerate mutase as a search model did not provide a satisfactory solution, indicating substantially different structures of these two phophoglycerate mutases.