Home LiteratureArticle Details
PMID: 21151115 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Crystal structure of the open conformation of the mammalian chaperonin CCT in complex with tubulin.

Nature structural & molecular biology ·Vol. 18 ·No. 1 ·2011-01-00 ·Pages 14-9

Muñoz IG, Yébenes H, Zhou M, Mesa P, Serna M, Park AY, Bragado-Nilsson E, Beloso A, de Cárcer G, Malumbres M, Robinson CV, Valpuesta JM, Montoya G

Abstract

Protein folding is assisted by molecular chaperones. CCT (chaperonin containing TCP-1, or TRiC) is a 1-MDa oligomer that is built by two rings comprising eight different 60-kDa subunits. This chaperonin regulates the folding of important proteins including actin, α-tubulin and β-tubulin. We used an electron density map at 5.5 Å resolution to reconstruct CCT, which showed a substrate in the inner cavities of both rings. Here we present the crystal structure of the open conformation of this nanomachine in complex with tubulin, providing information about the mechanism by which it aids tubulin folding. The structure showed that the substrate interacts with loops in the apical and equatorial domains of CCT. The organization of the ATP-binding pockets suggests that the substrate is stretched inside the cavity. Our data provide the basis for understanding the function of this chaperonin.

MeSH Terms
Animals Binding Sites Cattle Chaperonin Containing TCP-1/chemistry Crystallography, X-Ray Mass Spectrometry Models, Molecular Protein Folding Protein Structure, Tertiary Protein Subunits/chemistry Tubulin/chemistry,metabolism
Chemicals
Protein Subunits Tubulin Chaperonin Containing TCP-1
Authors & Affiliations
13 authors, click to expand affiliations / ORCID
Muñoz Inés G
Macromolecular Crystallography Group, Structural Biology and Biocomputing Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain.
Yébenes Hugo
Zhou Min
Mesa Pablo
Serna Marina
Park Ah Young
Bragado-Nilsson Elisabeth
Beloso Ana
de Cárcer Guillermo
Malumbres Marcos
Robinson Carol V
Valpuesta José M
Montoya Guillermo
References (43)
43 references, click to expand
  1. Closing the folding chamber of the eukaryotic chaperonin requires the transition state of ATP hydrolysis.
    Cell. 2003 May 2;113(3):369-81 PMID: 12732144
  2. Sequential ATP-induced allosteric transitions of the cytoplasmic chaperonin containing TCP-1 revealed by EM analysis.
    Nat Struct Mol Biol. 2005 Mar;12(3):233-7 PMID: 15696173
  3. Maximum likelihood refinement of electron microscopy data with normalization errors.
    J Struct Biol. 2009 May;166(2):234-40 PMID: 19236920
  4. Defining the eukaryotic cytosolic chaperonin-binding sites in human tubulins.
    J Mol Biol. 2000 Nov 17;304(1):81-98 PMID: 11071812
  5. The CCT chaperonin promotes activation of the anaphase-promoting complex through the generation of functional Cdc20.
    Mol Cell. 2003 Jul;12(1):87-100 PMID: 12887895
  6. Mechanism of lid closure in the eukaryotic chaperonin TRiC/CCT.
    Nat Struct Mol Biol. 2008 Jul;15(7):746-53 PMID: 18536725
  7. Role of the helical protrusion in the conformational change and molecular chaperone activity of the archaeal group II chaperonin.
    J Biol Chem. 2004 Apr 30;279(18):18834-9 PMID: 14978026
  8. NMR studies on the substrate-binding domains of the thermosome: structural plasticity in the protrusion region.
    J Mol Biol. 2004 Feb 20;336(3):717-29 PMID: 15095983
  9. Structure of bovine mitochondrial F(1)-ATPase inhibited by Mg(2+) ADP and aluminium fluoride.
    Structure. 2000 Jun 15;8(6):567-73 PMID: 10873854
  10. The 2.4 A crystal structure of the bacterial chaperonin GroEL complexed with ATP gamma S.
    Nat Struct Biol. 1996 Feb;3(2):170-7 PMID: 8564544
  11. Defining the TRiC/CCT interactome links chaperonin function to stabilization of newly made proteins with complex topologies.
    Nat Struct Mol Biol. 2008 Dec;15(12):1255-62 PMID: 19011634
  12. Function in protein folding of TRiC, a cytosolic ring complex containing TCP-1 and structurally related subunits.
    EMBO J. 1992 Dec;11(13):4767-78 PMID: 1361170
  13. Eukaryotic chaperonin CCT stabilizes actin and tubulin folding intermediates in open quasi-native conformations.
    EMBO J. 2000 Nov 15;19(22):5971-9 PMID: 11080144
  14. Structure of eukaryotic prefoldin and of its complexes with unfolded actin and the cytosolic chaperonin CCT.
    EMBO J. 2002 Dec 2;21(23):6377-86 PMID: 12456645
  15. XMIPP: a new generation of an open-source image processing package for electron microscopy.
    J Struct Biol. 2004 Nov;148(2):194-204 PMID: 15477099
  16. The unique hetero-oligomeric nature of the subunits in the catalytic cooperativity of the yeast Cct chaperonin complex.
    Proc Natl Acad Sci U S A. 1997 Sep 30;94(20):10780-5 PMID: 9380710
  17. 4.0-A resolution cryo-EM structure of the mammalian chaperonin TRiC/CCT reveals its unique subunit arrangement.
    Proc Natl Acad Sci U S A. 2010 Mar 16;107(11):4967-72 PMID: 20194787
  18. EMAN: semiautomated software for high-resolution single-particle reconstructions.
    J Struct Biol. 1999 Dec 1;128(1):82-97 PMID: 10600563
  19. Intracellular beta-tubulin/chaperonin containing TCP1-beta complex serves as a novel chemotherapeutic target against drug-resistant tumors.
    Cancer Res. 2009 Sep 1;69(17):6879-88 PMID: 19690144
  20. Cytosolic chaperonin protects folding intermediates of Gbeta from aggregation by recognizing hydrophobic beta-strands.
    Proc Natl Acad Sci U S A. 2006 May 30;103(22):8360-5 PMID: 16717193
  21. Essential function of the built-in lid in the allosteric regulation of eukaryotic and archaeal chaperonins.
    Nat Struct Mol Biol. 2007 May;14(5):432-40 PMID: 17460696
  22. Substructure solution with SHELXD.
    Acta Crystallogr D Biol Crystallogr. 2002 Oct;58(Pt 10 Pt 2):1772-9 PMID: 12351820
  23. The crystal structure of the bacterial chaperonin GroEL at 2.8 A.
    Nature. 1994 Oct 13;371(6498):578-86 PMID: 7935790
  24. Biochemical characterization of symmetric GroEL-GroES complexes. Evidence for a role in protein folding.
    J Biol Chem. 1996 Jan 5;271(1):68-76 PMID: 8550627
  25. Chaperonin complex with a newly folded protein encapsulated in the folding chamber.
    Nature. 2009 Jan 1;457(7225):107-10 PMID: 19122642
  26. Equivalent mutations in the eight subunits of the chaperonin CCT produce dramatically different cellular and gene expression phenotypes.
    J Mol Biol. 2010 Aug 20;401(3):532-43 PMID: 20600117
  27. Interpreting a low resolution map of human U1 snRNP using anomalous scatterers.
    Structure. 2009 Jul 15;17(7):930-8 PMID: 19604473
  28. Mechanism of the eukaryotic chaperonin: protein folding in the chamber of secrets.
    Trends Cell Biol. 2004 Nov;14(11):598-604 PMID: 15519848
  29. Improved methods for building protein models in electron density maps and the location of errors in these models.
    Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9 PMID: 2025413
  30. Substantial CCT activity is required for cell cycle progression and cytoskeletal organization in mammalian cells.
    Exp Cell Res. 2006 Jul 15;312(12):2309-24 PMID: 16765944
  31. Coot: model-building tools for molecular graphics.
    Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32 PMID: 15572765
  32. Crystal structure of the thermosome, the archaeal chaperonin and homolog of CCT.
    Cell. 1998 Apr 3;93(1):125-38 PMID: 9546398
  33. Maturation of human cyclin E requires the function of eukaryotic chaperonin CCT.
    Mol Cell Biol. 1998 Dec;18(12):7584-9 PMID: 9819444
  34. Mechanism of folding chamber closure in a group II chaperonin.
    Nature. 2010 Jan 21;463(7279):379-83 PMID: 20090755
  35. Methods used in the structure determination of bovine mitochondrial F1 ATPase.
    Acta Crystallogr D Biol Crystallogr. 1996 Jan 1;52(Pt 1):30-42 PMID: 15299723
  36. The inter-ring arrangement of the cytosolic chaperonin CCT.
    EMBO Rep. 2007 Mar;8(3):252-7 PMID: 17304242
  37. A cytoplasmic chaperonin that catalyzes beta-actin folding.
    Cell. 1992 Jun 12;69(6):1043-50 PMID: 1351421
  38. Cytoplasmic chaperonin containing TCP-1: structural and functional characterization.
    Biochemistry. 1997 May 13;36(19):5817-26 PMID: 9153422
  39. Eukaryotic type II chaperonin CCT interacts with actin through specific subunits.
    Nature. 1999 Dec 9;402(6762):693-6 PMID: 10604479
  40. CCT chaperonin complex is required for the biogenesis of functional Plk1.
    Mol Cell Biol. 2005 Jun;25(12):4993-5010 PMID: 15923617
  41. Domain-specific chaperone-induced expansion is required for beta-actin folding: a comparison of beta-actin conformations upon interactions with GroEL and tail-less complex polypeptide 1 ring complex (TRiC).
    Biochemistry. 2007 Nov 6;46(44):12639-47 PMID: 17939680
  42. Placement of protein and RNA structures into a 5 A-resolution map of the 50S ribosomal subunit.
    Nature. 1999 Aug 26;400(6747):841-7 PMID: 10476961
  43. Elucidation of the subunit orientation in CCT (chaperonin containing TCP1) from the subunit composition of CCT micro-complexes.
    EMBO J. 1997 Jul 16;16(14):4311-6 PMID: 9250675
Article Info
Journal
Nature structural & molecular biology
Abbr.
Nat Struct Mol Biol
ISSN
1545-9985
DOI
10.1038/nsmb.1971
Published
2011-01-00
Epub
2010-00-12
Pages
14-9
Language
English
Region
United States
NLM ID
101186374
Subset
IM
Databases
PDB
2XSM
External Links
PubMed source DOI: 10.1038/nsmb.1971 View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2