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Purification, crystallization, and preliminary X-ray crystallographic analysis of the Group III chaperonin from Carboxydothermus hydrogenoformans
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Purification, crystallization, and preliminary X-ray crystallographic analysis of the Group III chaperonin from Carboxydothermus hydrogenoformans
Young Jun An 1, Sara E. Rowland 2, Frank T. Robb 2,3, Sun-Shin Cha 4
Journal of Microbiology 2016;54(6):440-444
DOI: https://doi.org/10.1007/s12275-016-6089-5
Published online: May 27, 2016
1Korea Institute of Ocean Science and Technology, Ansan 15627, Republic of Korea, 2Institute of Marine and Environmental Technology, Baltimore, MD 21202, and Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA, 3Department of Microbiology and Immunology, University of Maryland, Baltimore, MD, 21202, USA, 4Department of Chemistry and Nano Science, Ewha Womans University, Seoul 03760, Republic of Korea1Korea Institute of Ocean Science and Technology, Ansan 15627, Republic of Korea, 2Institute of Marine and Environmental Technology, Baltimore, MD 21202, and Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA, 3Department of Microbiology and Immunology, University of Maryland, Baltimore, MD, 21202, USA, 4Department of Chemistry and Nano Science, Ewha Womans University, Seoul 03760, Republic of Korea
Corresponding author:  Sun-Shin Cha , Tel: +82-2-3277-3420, 
Received: 29 February 2016   • Revised: 6 April 2016   • Accepted: 18 April 2016
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Chaperonins (CPNs) are megadalton sized ATP-dependent nanomachines that facilitate protein folding through complex cycles of complex allosteric articulation. They consist of two back-to-back stacked multisubunit rings. CPNs are usually classified into Group I and Group II. Here, we report the crystallization of both the AMPPNP (an ATP analogue) and ADP bound forms of a novel CPN, classified as belonging to a third Group, recently discovered in the extreme thermophile Carboxydothermus hydrogenoformans. Crystals of the two forms were grown by the vapor batch crystallization
method
at 295 K. Crystals of the Ch-CPN/AMPPNP complex diffracted to 3.0 Å resolution and belonged to the space group P422, with unit-cell parameters a = b = 186.166, c = 160.742 Å. Assuming the presence of four molecules in the asymmetric unit, the solvent content was estimated to be about 60.02%. Crystals of the Ch-CPN/ADP complex diffracted to 4.0 Å resolution and belonged to the space group P4212, with unit-cell parameters a = b = 209.780, c = 169.813Å. Assuming the presence of four molecules in the asymmetric unit, the solvent content was estimated to be about 70.19%.

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    Purification, crystallization, and preliminary X-ray crystallographic analysis of the Group III chaperonin from Carboxydothermus hydrogenoformans
    J. Microbiol. 2016;54(6):440-444.   Published online May 27, 2016
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