title: Purification, crystallization, and preliminary X-ray crystallographic analysis of the Group III chaperonin from Carboxydothermus hydrogenoformans.
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%.
authors.affiliation: Korea Institute of Ocean Science and Technology, Ansan, 15627, Republic of Korea.
authors.forename: Young Jun
authors.initials: YJ
authors.lastname: An
authors.affiliation: Institute of Marine and Environmental Technology, Baltimore, MD, 21202, USA.|Institute for Bioscience and Biotechnology Research, Rockville, MD, 20850, USA.
authors.forename: Sara E
authors.initials: SE
authors.lastname: Rowland
authors.affiliation: Institute of Marine and Environmental Technology, Baltimore, MD, 21202, USA.|Institute for Bioscience and Biotechnology Research, Rockville, MD, 20850, USA.|Department of Microbiology and Immunology, University of Maryland, Baltimore, MD, 21202, USA.
authors.forename: Frank T
authors.initials: FT
authors.lastname: Robb
authors.affiliation: Department of Chemistry and Nano Science, Ewha Womans University, Seoul, 03760, Republic of Korea. chajung@ewha.ac.kr.
authors.forename: Sun-Shin
authors.initials: SS
authors.lastname: Cha
mesh_terms: D000244:Adenosine Diphosphate / Q000737:chemistry / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D018833:Chaperonins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D046950:Thermoanaerobacterium / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D018833:Chaperonins
keywords: AT-Pfueled nanomachine; Group III chaperonins; crystallization; protein folding