Cytosolic chaperonin CCT possesses GTPase activity

DOI: 10.4236/ajmb.2011.13013   PDF   HTML   XML   4,677 Downloads   10,091 Views   Citations


Cytosolic chaperonin CCT (also known as TRiC) is a hetero-oligomeric cage-like molecular chaperone that assists in protein folding by ATPase cycle-dependent conformational changes. However, role of the nucleo-tide binding and hydrolysis in CCT-assisted protein folding is still poorly understood. We purified CCT by using ATP-Sepharose and other columns, and found that CCT possesses ability to hydrolyze GTP, with an activity level very similar to the ATPase activity. CCT was more resistant to proteinase K treatment in the presence of GTP or ATP. These results suggest that the GTPase activity of CCT may play a role in chaperone-assisted protein folding.

Share and Cite:

Noguchi, S. , Toyoshima, K. , Yamamoto, S. , Miyazaki, T. , Otaka, M. , Watanabe, S. , Imai, K. , Senoo, H. , Kobayashi, R. , Jikei, M. , Kawata, Y. , Kubota, H. and Itoh, H. (2011) Cytosolic chaperonin CCT possesses GTPase activity. American Journal of Molecular Biology, 1, 123-130. doi: 10.4236/ajmb.2011.13013.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Bukau, B. and Horwich, A.L. (1998) The Hsp70 and Hsp60 chaperone machines. Cell, 14, 351-366. HHdoi:10.1016/S0092-8674(00)80928-9
[2] Hartl, F.U. and Hayer-Hartl, M. (2002) Molecular cha-perones in the cytosol: From nascent chain to folded protein. Science, 295, 1852-1858. HHdoi:10.1126/science.1068408
[3] Horwich, A.L., Fenton, W.A., Chapman E. and Farr, G.W. (2007) Two Families of Chaperonin: Physiology and Mechanism. Annual Review of Cell and Developmental Biology, 23, 115-145. HHdoi:10.1146/annurev.cellbio.23.090506.123555
[4] Itoh, H., Kobayashi, R., Wakui, H., Komatsuda, A., Ohtani, H., Miura, A.B., Otaka, M., Masamune, O., Andoh, H., Koyama, K., Sato, Y. and Tashima, Y. (1995) Mammalian 60-kDa stress protein (chaperonin homolog). Identification, biochemical properties, and localization. Journal of Biological Chemistry, 270, 13429-1335.
[5] Itoh H, Komatsuda A, Wakui H, Miura AB, Tashima Y. (1999) Mammalian HSP60 is a major target for an immunosuppressant mizoribine. Journal of Biological Chemistry, 274, 35147-35151. HHdoi:10.1074/jbc.274.49.35147
[6] Itoh H, Komatsuda A, Ohtani H, Wakui H, Imai H, Sawada K, Otaka M, Ogura M, Suzuki A, Hamada F. (2002) Mammalian HSP60 is quickly sorted into the mitochondria under conditions of dehydration. European Journal of Biochemical, 269, 5931-5938. HHdoi:10.1046/j.1432-1033.2002.03317.x
[7] Gutsche, I., Essen, L.O. and Baumeister, W. (1999) Group II chaperonins: New TRiC(k)s and turns of a protein folding machine. Journal of Molecular Biology, 2-93, 295-312. HHdoi:10.1006/jmbi.1999.3008
[8] Spiess, C., Meyer, A.S., Reissmann, S. and Frydman, J. (2004) Mechanism of the eukaryotic chaperonin: Protein folding in the chamber of secrets. Trends Cell Biology, 14, 598-604. HHdoi:10.1016/j.tcb.2004.09.015
[9] Sigler, P.B., Xu, Z., Rye, H.S., Burston, S.G., Fenton, W.A. and Horwich, A.L. (1999) Structure and function in GroEL-mediated protein folding. Annual Review of Biochemistry, 67, 581-608. doi:10.1146/annurev.biochem.67.1.581
[10] Iizuka, R., So, S., Inobe, T., Yoshida, T., Zako, T., Kuwajima, K. and Yohda, M. (2004) Role of the helical protrusion in the conformational change and molecular chaperone activity of the archaeal group II chaperonin. Journal of Biology Chemistry, 279, 18834-18839.
[11] Kubota, H. 2002. Function and regulation of cytosolic molecular chaperone CCT. Vitam Horm, 65, 313-331. doi:10.1016/S0083-6729(02)65069-1
[12] Kubota, H., Hynes, G., Carne, A., Ashworth, A. and Willison, K. (1994) Identification of six Tcp-1-related genes encoding divergent subunits of the TCP-1-containing chaperonin. Current Biology, 4, 89-99. doi:10.1016/S0960-9822(94)00024-2
[13] Frydman, J., Nimmesgern, E., Ohtuka, K. and Hartl, F.-U. (1994) Folding of nascent polypeptide chains in a high molecular mass assembly with molecular chaperones. Nature, 370, 111-117. doi:10.1038/370111a0
[14] Tian, G., Vainberg, I.E., Tap, W.D., Lewis, S.A. and Cowan, N.J. (1995) Specificity in chaperonin-mediated protein folding. Nature, 375, 250-253. doi:10.1038/375250a0
[15] Kubota, S., Kubota, H. and Nagata, K. (2006) Cytosolic chaperonin protects folding intermediates of Gbeta from aggregation by recognizing hydrophobic beta-strands. Proceedings of the National Academy of Sciences, USA, 103, 8360-8365. doi:10.1073/pnas.0600195103
[16] Llorca, O., Martin-Benito, J., Grantham, J., Ritco-Vonsovici, M., Willison, K.R., Carrascosa, J.L. and Valpuesta, J.M. (2001) The "sequential allosteric ring" mechanism in the eukaryotic chaperonin-assisted folding of actin and tubulin. EMBO Journal, 20, 4065-4075. doi:10.1093/emboj/20.15.4065
[17] Meyer, A.S., Gillespie, J.R., Walther, D., Millet, I.S., Doniach, S. and Frydman, J. (2003) Closing the folding chamber of the eukaryotic chaperonin requires the transition state of ATP hydrolysis. Cell, 113, 369-381. doi:10.1016/S0092-8674(03)00307-6
[18] Lin, P. and Sherman, F. (1997) The unique hetero-oligomeric nature of the subunits in the catalytic cooperativity of the yeast Cct chaperonin complex. Proceedings of the National Academy of Sciences, USA, 94, 10780-10785. doi:10.1073/pnas.94.20.10780
[19] Rivenzon-Segal, D., Wolf, S.G., Shimon, L., Willison. K.R. and Horovitz, A. (2005) Sequential ATP-induced allosteric transitions of the cytoplasmic chaperonin containing TCP-1 revealed by EM analysis. Nature Structural and Molecular Biologlogy, 12, 233-237. doi:10.1038/nsmb901
[20] Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685. doi:10.1038/227680a0
[21] Towbin, H., Staehelin, T. and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proceedings of the National Academy of Sciences, USA, 76, 4350-4354. doi:10.1073/pnas.76.9.4350
[22] O'Farrell, P.H. (1975) High resolution two-dimensional electrophoresis of proteins. Journal of Biological Chemistry, 250, 4007-4021.
[23] Martin, B., Pallen, C.J., Wang, J.H. and Graves, D.J. (1985) Use of fluorinated tyrosine phosphates to probe the substrate specificity of the low molecular weight phosphatase activity of calcineurin. Journal of Biological Chemistry, 260, 14932-14937.
[24] Lake, J.A. (1978) Electron microscopy of specific proteins: Three-dimensional mapping of ribosomal proteins using antibody labels. In: Koehler, J.K., Eds., Advanced techniques in biological electron microscopy, Springer-Verlag, New York, 2, 173-211.
[25] Liou, A.K. and Willison, K.R. (1997) Elucidation of the subunit orientation in CCT (chaperonin containing TCP1) from the subunit composition of CCT micro-complexes. EMBO Journal, 16, 4311-4316. doi:10.1093/emboj/16.14.4311
[26] Douglas, N.R., Reissmann, S., Zhang, J., Chen, B., Ja-kana, J., Kumar, R., Chiu, W. and Frydman, J. (2011) Dual action of ATP hydrolysis couples lid closure to substrate release into the group II chaperonin chamber. Cell, 144, 240-252. doi:10.1016/j.cell.2010.12.017
[27] Hirai, H., Noi, K., Hongo, K., Mizobata, T. and Kawata, Y. (2008) Functional characterization of the recombinant group II chaperonin alpha from thermoplasma acidophilum. Journal of Biochemistry, 143, 505-515. doi:10.1093/jb/mvm241

comments powered by Disqus

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.