| Title | Purification, crystallization, and preliminary X-ray crystallographic analysis of the Group III chaperonin from Carboxydothermus hydrogenoformans |
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| Author | Young Jun An1, Sara E. Rowland2, Frank T. Robb2,3, and Sun-Shin Cha4* |
| Address | 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 Korea |
| Bibliography | Journal of Microbiology, 54(6),440-444, 2016, |
| DOI | 10.1007/s12275-016-6089-5 |
| Key Words | Group III chaperonins, protein folding, ATPfueled nanomachine, crystallization |
| Abstract | 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%. |