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

详细信息    查看全文

• 作者：Xinghua Qin ; Ziwei Chen ; Ping Li and Guoqin Liu
• 关键词：plant-specific kinesin ; CH domain ; cotton kinesin GhKCH2
• 刊名：Acta Crystallographica Section F
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：72
• 期：3
• 页码：240-243
• 全文大小：363K
• ISSN：1744-3091

文摘

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 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, α = 90.00, β = 97.31, γ = 90.00°. Four molecules were found in the asymmetric unit with a Matthews coefficient of 2.22 ų Da⁻¹, corresponding to a solvent content of 44.8%.