Open AccessCloning, purification, crystallization and preliminary X‐ray analysis of the receiver domain of the histidine kinase CKI1 from Arabidopsis thaliana
Author(s) -
Klumpler Tomáš,
Pekárová Blanka,
Marek Jaromír,
Borkovcová Petra,
Janda Lubomír,
Hejátko Jan
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109012032
Subject(s) - crystallography , arabidopsis thaliana , mosaicity , molecular replacement , chemistry , crystallization , ammonium sulfate , histidine , x ray crystallography , materials science , diffraction , chromatography , biochemistry , crystal structure , optics , physics , organic chemistry , amino acid , mutant , gene
The receiver domain (RD) of a sensor histidine kinase (HK) catalyses the transphosphorylation reaction during the action of HKs in hormonal and abiotic signalling in plants. Crystals of the recombinant RD of the Arabidopsis thaliana HK CYTOKININ‐INDEPENDENT1 (CKI1 RD ) have been obtained by the hanging‐drop vapour‐diffusion method using ammonium sulfate as a precipitant and glycerol as a cryoprotectant. The crystals diffracted to approximately 2.4 Å resolution on beamline BW7B of the DORIS‐III storage ring. The diffraction improved significantly after the use of a non‐aqueous cryoprotectant. Crystals soaked in Paratone‐N diffracted to at least 2.0 Å resolution on beamline BW7B and their mosaicity decreased more than tenfold. The crystals belonged to space group C 222 1 , with unit‐cell parameters a = 54.46, b = 99.82, c = 79.94 Å. Assuming the presence of one molecule of the protein in the asymmetric unit gives a Matthews coefficient V M of 2.33 Å 3 Da −1 . A molecular‐replacement solution has been obtained and structure refinement is in progress.