Open AccessCrystallization and preliminary X‐ray analysis of the stress‐response PPM phosphatase RsbX from Bacillus subtilis
Author(s) -
Suganuma Masatoshi,
Teh Aik Hong,
Makino Masatomo,
Shimizu Nobutaka,
Kaneko Tomonori,
Hirata Kunio,
Yamamoto Masaki,
Kumasaka Takashi
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109038846
Subject(s) - bacillus subtilis , crystallization , crystallography , escherichia coli , phosphatase , dephosphorylation , triclinic crystal system , rna polymerase , chemistry , x ray crystallography , biophysics , diffraction , biology , enzyme , crystal structure , biochemistry , bacteria , gene , physics , optics , genetics , organic chemistry
RsbX from Bacillus subtilis is a manganese‐dependent PPM phosphatase and negatively regulates the signal transduction of the general stress response by the dephosphorylation of RsbS and RsbR, which are activators of the alternative RNA polymerase σ factor SigB. In order to elucidate the structural–functional relationship of its Ser/Thr protein‐phosphorylation mechanism, an X‐ray crystallographic diffraction study of RsbX was performed. Recombinant RsbX was expressed in Escherichia coli , purified and crystallized. Crystals were obtained using the sitting‐drop vapour‐diffusion method and X‐ray diffraction data were collected to 1.06 Å resolution with an R merge of 8.1%. The crystals belonged to the triclinic space group P 1, with unit‐cell parameters a = 33.3, b = 41.7, c = 68.6 Å, α = 98.8, β = 90.0, γ = 108.4°.