Crystallization, characterization and preliminary X-ray crystallographic analysis of GK2848, a putative carbonic anhydrase of Geobacillus kaustophilus.
作者： Preethi Ragunathan , Gokul Raghunath , Seiki Kuramitsu , Shigeyuki Yokoyama , Thirumananseri Kumarevel
DOI: 10.1107/S1744309112051913
关键词:
摘要: GK2848, a hypothetical protein from the thermophilic organism Geobacillus kaustophilus, was cloned and overexpressed in Escherichia coli. The purified to homogeneity using Ni–NTA affinity-column gel-filtration chromatography. crystallized sitting-drop vapour-diffusion method. crystals diffracted resolution of 2.70 A belonged orthorhombic space group P21212. GK2848 bears sequence homology carbonic anhydrases various bacterial species, indicating that it belongs anhydrase family proteins. A subsequent activity assay Wilbur–Anderson method confirmed its function as anhydrase. preliminary structure solution obtained by molecular replacement MOLREP. Mutation biochemical characterization are progress. functional analysis might provide valuable information on novel class anhydrases, none homologous structures have been characterized.
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