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Expression, purification, crystallization and preliminary X‐ray analysis of the KaiC‐like protein PH0187 from the hyperthermophilic archaeon Pyrococcus horikoshii OT3
Author(s) -
Kang HeeJin,
Kubota Keiko,
Miyazono Kenichi,
Tanokura Masaru
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110048426
Subject(s) - pyrococcus horikoshii , crystallization , materials science , crystallography , chemistry , crystal structure , organic chemistry
KaiC is the central protein in the circadian rhythm in cyanobacteria. The 28 kDa KaiC‐like protein PH0187 from the hyperthermophilic archaeon Pyrococcus horikoshii was expressed in Escherichia coli , purified and crystallized using the sitting‐drop vapour‐diffusion method at 293 K. Crystals of PH0187 were obtained using a reservoir solution consisting of 1.0  M ammonium phosphate monobasic and 0.1  M sodium citrate tribasic pH 5.3 (the final pH value of the reservoir solution was 4.8) and diffracted X‐rays to 2.75 Å resolution. The crystal of PH0187 belonged to space group P 6 3 22, with unit‐cell parameters a  =  b  = 239.1, c = 106.5 Å. The crystal contained four PH0187 molecules in the asymmetric unit.

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