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- Purification, crystallization, and preliminary X-ray crystallographic analysis of the Group III chaperonin from Carboxydothermus hydrogenoformans
- Young Jun An , Sara E. Rowland , Frank T. Robb , Sun-Shin Cha
- J. Microbiol. 2016;54(6):440-444. Published online May 27, 2016
- DOI: https://doi.org/10.1007/s12275-016-6089-5
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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%. -
Citations
Citations to this article as recorded by
- Bridging human chaperonopathies and microbial chaperonins
Everly Conway de Macario, Masafumi Yohda, Alberto J. L. Macario, Frank T. Robb
Communications Biology.2019;[Epub] CrossRef - Prokaryotic Chaperonins as Experimental Models for Elucidating Structure-Function Abnormalities of Human Pathogenic Mutant Counterparts
Everly Conway de Macario, Frank T. Robb, Alberto J. L. Macario
Frontiers in Molecular Biosciences.2017;[Epub] CrossRef - Structural and mechanistic characterization of an archaeal-like chaperonin from a thermophilic bacterium
Young Jun An, Sara E. Rowland, Jung-Hyun Na, Dario Spigolon, Seung Kon Hong, Yeo Joon Yoon, Jung-Hyun Lee, Frank T. Robb, Sun-Shin Cha
Nature Communications.2017;[Epub] CrossRef
- Bridging human chaperonopathies and microbial chaperonins
Research Support, Non-U.S. Gov't
- Experimental Phasing Using Zinc and Sulfur Anomalous Signals Measured at the Zinc Absorption Peak
- Sangmin Lee , Min-Kyu Kim , Chang-Jun Ji , Jin-Won Lee , Sun-Shin Cha
- J. Microbiol. 2013;51(5):639-643. Published online October 31, 2013
- DOI: https://doi.org/10.1007/s12275-013-3412-2
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Abstract
- Iron is an essential transition metal required for bacterial growth and survival. Excess free iron can lead to the generation of reactive oxygen species that can cause severe damage to cellular functions. Cells have developed iron-sensing regulators to maintain iron homeostasis at the transcription level. The ferric uptake regulator (Fur) is an iron-responsive regulator that controls the expression of genes involved in iron homeostasis, bacterial virulence, stress resistance, and redox metabolism. Here, we report the expression, purification, crystallization, and phasing of the apo-form of Bacillus subtilis Fur (BsFur) in the absence of regulatory metal ions. Crystals were obtained by microbatch crystallization method at 295 K and diffraction data at a resolution of 2.6 Å was collected at the zinc peak wavelength (λ=1.2823 Å). Experimental phasing identified the positions of one zinc atom and four sulfur atoms of cysteine residues coordinating the zinc atom, indicating that the data contained a meaningful anomalous scattering originating from the ordered zinc-coordinating sulfur atoms, in spite of the small anomalous signals of sulfur atoms at the examined wavelength.
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