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Crystallization and X‐ray diffraction analysis of the CH domain of the cotton kinesin GhKCH2
Author(s) -
Qin Xinghua,
Chen Ziwei,
Li Ping,
Liu Guoqin
Publication year - 2016
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x16001825
Subject(s) - crystallography , crystallization , chemistry , kinesin , tris , molecule , solvent , calponin , peg ratio , x ray crystallography , stereochemistry , diffraction , actin , biology , biochemistry , physics , organic chemistry , microtubule , finance , economics , microbiology and biotechnology , optics
GhKCH2 belongs to a group of plant‐specific kinesins (KCHs) containing an actin‐binding calponin homology (CH) domain in the N‐terminus. Previous studies revealed that the GhKCH2 CH domain (GhKCH2‐CH) had a higher affinity for F‐actin ( K d = 0.42 ± 0.02 µ M ) than most other CH‐domain‐containing proteins. To understand the underlying mechanism, prokaryotically expressed GhKCH2‐CH (amino acids 30–166) was purified and crystallized. Crystals were grown by the sitting‐drop vapour‐diffusion method using 0.1 M Tris–HCl pH 7.0, 20%( w / v ) PEG 8000 as a precipitant. The crystals diffracted to a resolution of 2.5 Å and belonged to space group P 2 1 , with unit‐cell parameters a = 41.57, b = 81.92, c = 83.00 Å, α = 90.00, β = 97.31, γ = 90.00°. Four molecules were found in the asymmetric unit with a Matthews coefficient of 2.22 Å 3 Da −1 , corresponding to a solvent content of 44.8%.