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%.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Balch, W.E., Morimoto, R.I., Dillin, A., and Kelly, J.W. 2008. Adapting proteostasis for disease intervention. Science 319, 916–919.
Bigotti, M.G. and Clarke, A.R. 2008. Chaperonins: The hunt for the group II mechanism. Arch. Biochem. Biophys. 474, 331–339.
Bukau, B. and Horwich, A.L. 1998. The hsp70 and hsp60 chaperone machines. Cell 92, 351–366.
Collaborative Computational Project, N. 1994. The ccp4 suite: Programs for protein crystallography. Acta Crystallogr. D Biol. Crystallogr. 50, 760–763.
Ditzel, L., Lowe, J., Stock, D., Stetter, K.O., Huber, H., Huber, R., and Steinbacher, S. 1998. Crystal structure of the thermosome, the archaeal chaperonin and homolog of CCT. Cell 93, 125–138.
Frydman, J., Nimmesgern, E., Erdjument-Bromage, H., Wall, J.S., Tempst, P., and Hartl, F.U. 1992. Function in protein folding of TRiC, a cytosolic ring complex containing TCP-1 and structurally related subunits. EMBO J. 11, 4767–4778.
Gao, Y., Thomas, J.O., Chow, R.L., Lee, G.H., and Cowan, N.J. 1992. A cytoplasmic chaperonin that catalyzes β-actin folding. Cell 69, 1043–1050.
Kabsch, W. 2010. XDS. Acta Crystallogr. D Biol. Crystallogr. 66, 125–132.
Kim, S., Willison, K.R., and Horwich, A.L. 1994. Cystosolic chaperonin subunits have a conserved atpase domain but diverged polypeptide-binding domains. Trends Biochem. Sci. 19, 543–548.
Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin, F., Wallace, I.M., Wilm, A., Lopez, R., et al. 2007. Clustal W and Clustal X version 2.0. Bioinformatics 23, 2947–2948.
Otwinowski, Z. and Minor, W. 1997. Processing of X-ray diffraction data collected in oscillation mode. Method. Enzymol. 276, 307–326.
Pereira, J.H., Ralston, C.Y., Douglas, N.R., Meyer, D., Knee, K.M., Goulet, D.R., King, J.A., Frydman, J., and Adams, P.D. 2010. Crystal structures of a group II chaperonin reveal the open and closed states associated with the protein folding cycle. J. Biol. Chem. 285, 27958–27966.
Phipps, B.M., Hoffmann, A., Stetter, K.O., and Baumeister, W. 1991. A novel ATPase complex selectively accumulated upon heat shock is a major cellular component of thermophilic archaebacteria. EMBO J. 10, 1711–1722.
Ranson, N.A., Clare, D.K., Farr, G.W., Houldershaw, D., Horwich, A.L., and Saibil, H.R. 2006. Allosteric signaling of ATP hydrolysis in GroEL-GroES complexes. Nat. Struct. Mol. Biol. 13, 147–152.
Techtmann, S.M., Colman, A.S., Murphy, M.B., Schackwitz, W.S., Goodwin, L.A., and Robb, F.T. 2011. Regulation of multiple carbon monoxide consumption pathways in anaerobic bacteria. Front. Microbiol. 2, 147.
Techtmann, S.M., Lebedinsky, A.V., Colman, A.S., Sokolova, T.G., Woyke, T., Goodwin, L., and Robb, F.T. 2012. Evidence for horizontal gene transfer of anaerobic carbon monoxide dehydrogenases. Front. Microbiol. 3, 132.
Techtmann, S.M. and Robb, F.T. 2010. Archaeal-like chaperonins in bacteria. Proc. Natl. Acad. Sci. USA 107, 20269–20274.
Tilly, K., Murialdo, H., and Georgopoulos, C. 1981. Identification of a second Escherichia coli groE gene whose product is necessary for bacteriophage morphogenesis. Proc. Natl. Acad. Sci. USA 78, 1629–1633.
Wu, M., Ren, Q., Durkin, A.S., Daugherty, S.C., Brinkac, L.M., Dodson, R.J., Madupu, R., Sullivan, S.A., Kolonay, J.F., Haft, D.H., et al. 2005. Life in hot carbon monoxide: The complete genome sequence of Carboxydothermus hydrogenoformans Z-2901. PLoS Genet. 1, e65.
Xu, Z., Horwich, A.L., and Sigler, P.B. 1997. The crystal structure of the asymmetric GroEL-GroES-(ADP)7 chaperonin complex. Nature 388, 741–750.
Yaffe, M.B., Farr, G.W., Miklos, D., Horwich, A.L., Sternlicht, M.L., and Sternlicht, H. 1992. TCP1 complex is a molecular chaperone in tubulin biogenesis. Nature 358, 245–248.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
An, Y.J., Rowland, S.E., Robb, F.T. et al. Purification, crystallization, and preliminary X-ray crystallographic analysis of the Group III chaperonin from Carboxydothermus hydrogenoformans . J Microbiol. 54, 440–444 (2016). https://doi.org/10.1007/s12275-016-6089-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-016-6089-5