Crystallization and preliminary X-ray crystallographic analysis of the β-N-acetylglucosaminidase CbsA from Thermotoga neapolitana.

Bo-Young Yoon, Li Jiao, Hyung Ryong Moon, Jaeho Cha, Nam-Chul Ha

College of Pharmacy and Research Institute for Drug Development, Pusan National University, Busan 609-735, Republic of Korea.

Acta crystallographica. Section F, Structural biology and crystallization communications 2012 Jan 1

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The β-N-acetylglucosaminidase CbsA was cloned from the thermophilic Gram-negative bacterium Thermotoga neapolitana. Although CbsA contains a family 3 glycoside hydrolase-type (GH3-type) catalytic domain, it can be distinguished from other GH3-type β-N-acetylglucosaminidases by its high activity towards chitobiose. The homodimeric CbsA contains a unique domain at the C-terminus for which the three-dimensional structure is not yet known. In this study, CbsA was overexpressed and the recombinant protein was purified using Ni-NTA affinity and gel-filtration chromatography. The purified CbsA protein was crystallized using the vapour-diffusion method. A diffraction data set was collected to a resolution of 2.0 Å at 100 K. The crystal belonged to space group R32. To obtain initial phases, the crystallization of selenomethionyl-substituted protein and the production of heavy-atom derivative crystals are in progress. © 2012 International Union of Crystallography. All rights reserved.

Citation

Bo-Young Yoon, Li Jiao, Hyung Ryong Moon, Jaeho Cha, Nam-Chul Ha. Crystallization and preliminary X-ray crystallographic analysis of the β-N-acetylglucosaminidase CbsA from Thermotoga neapolitana. Acta crystallographica. Section F, Structural biology and crystallization communications. 2012 Jan 1;68(Pt 1):56-8