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Crystallization and X-ray diffraction analysis of the CH domain of the cotton kinesin GhKCH2.

ABSTRACT: 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 (Kd = 0.42 ± 0.02 µM) 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 P21, with unit-cell parameters a = 41.57, b = 81.92, c = 83.00 Å, ? = 90.00, ? = 97.31, ? = 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%.

SUBMITTER: Qin X

PROVIDER: S-EPMC4774884 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature

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Crystallization and X-ray diffraction analysis of the CH domain of the cotton kinesin GhKCH2.

Qin Xinghua X Chen Ziwei Z Li Ping P Liu Guoqin G

Acta crystallographica. Section F, Structural biology communications 20160219 Pt 3

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 (Kd = 0.42 ± 0.02 µM) 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 usin ...[more]

PMID: 26919529

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