title: Crystallization, characterization and preliminary X-ray crystallographic analysis of GK2848, a putative carbonic anhydrase of Geobacillus kaustophilus.
abstract: GK2848, a hypothetical protein from the thermophilic organism Geobacillus kaustophilus, was cloned and overexpressed in Escherichia coli. The protein was purified to homogeneity using Ni-NTA affinity-column and gel-filtration chromatography. The purified protein was crystallized using the sitting-drop vapour-diffusion method. The crystals diffracted to a resolution of 2.70 Å and belonged to the orthorhombic space group P2(1)2(1)2. GK2848 bears sequence homology to carbonic anhydrases of various bacterial species, indicating that it belongs to the carbonic anhydrase family of proteins. A subsequent carbonic anhydrase activity assay of GK2848 using the Wilbur-Anderson method confirmed its function as a carbonic anhydrase. A preliminary structure solution was obtained by molecular replacement using MOLREP. Mutation and biochemical characterization of the protein are in progress. The structure and functional analysis of GK2848 might provide valuable information on a novel class of carbonic anhydrases, as none of its homologous structures have been characterized.
authors: Centre of Advanced Study in Crystallography and Biophysics, University of Madras, Guindy Campus, Chennai 600 025, India.
authors: Preethi
authors: P
authors: Ragunathan
authors: Gokul
authors: G
authors: Raghunath
authors: Seiki
authors: S
authors: Kuramitsu
authors: Shigeyuki
authors: S
authors: Yokoyama
authors: Thirumananseri
authors: T
authors: Kumarevel
authors: Karthe
authors: K
authors: Ponnuraj
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D002256:Carbonic Anhydrases / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D056506:Geobacillus / Q000201:enzymology*; D008969:Molecular Sequence Data; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D002256:Carbonic Anhydrases
keywords: Geobacillus kaustophilus; carbonic anhydrases; hypothetical proteins; left-handed β-helix
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