The molecular architecture of the eukaryotic chaperonin TRiC/CCT

Alexander Leitner; Lukasz A. Joachimiak; Andreas Bracher; Leonie Mönkemeyer; Thomas Walzthoeni; Bryan Chen; Sebastian Pechmann; Susan Holmes; Yao Cong; Boxue Ma; Steve Ludtke; Wah Chiu; F. Ulrich Hartl; Ruedi Aebersold; Judith Frydman

doi:10.1016/j.str.2012.03.007

The molecular architecture of the eukaryotic chaperonin TRiC/CCT

Alexander Leitner, Lukasz A. Joachimiak, Andreas Bracher, Leonie Mönkemeyer, Thomas Walzthoeni, Bryan Chen, Sebastian Pechmann, Susan Holmes, Yao Cong, Boxue Ma, Steve Ludtke, Wah Chiu, F. Ulrich Hartl, Ruedi Aebersold, Judith Frydman

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233 Scopus citations

Abstract

TRiC/CCT is a highly conserved and essential chaperonin that uses ATP cycling to facilitate folding of approximately 10% of the eukaryotic proteome. This 1 MDa hetero-oligomeric complex consists of two stacked rings of eight paralogous subunits each. Previously proposed TRiC models differ substantially in their subunit arrangements and ring register. Here, we integrate chemical crosslinking, mass spectrometry, and combinatorial modeling to reveal the definitive subunit arrangement of TRiC. In vivo disulfide mapping provided additional validation for the crosslinking-derived arrangement as the definitive TRiC topology. This subunit arrangement allowed the refinement of a structural model using existing X-ray diffraction data. The structure described here explains all available crosslink experiments, provides a rationale for previously unexplained structural features, and reveals a surprising asymmetry of charges within the chaperonin folding chamber.

Original language	English (US)
Pages (from-to)	814-825
Number of pages	12
Journal	Structure
Volume	20
Issue number	5
DOIs	https://doi.org/10.1016/j.str.2012.03.007
State	Published - May 9 2012
Externally published	Yes

ASJC Scopus subject areas

Structural Biology
Molecular Biology

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10.1016/j.str.2012.03.007

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@article{fded949255af429da1824a0501b2595b,

title = "The molecular architecture of the eukaryotic chaperonin TRiC/CCT",

abstract = "TRiC/CCT is a highly conserved and essential chaperonin that uses ATP cycling to facilitate folding of approximately 10% of the eukaryotic proteome. This 1 MDa hetero-oligomeric complex consists of two stacked rings of eight paralogous subunits each. Previously proposed TRiC models differ substantially in their subunit arrangements and ring register. Here, we integrate chemical crosslinking, mass spectrometry, and combinatorial modeling to reveal the definitive subunit arrangement of TRiC. In vivo disulfide mapping provided additional validation for the crosslinking-derived arrangement as the definitive TRiC topology. This subunit arrangement allowed the refinement of a structural model using existing X-ray diffraction data. The structure described here explains all available crosslink experiments, provides a rationale for previously unexplained structural features, and reveals a surprising asymmetry of charges within the chaperonin folding chamber.",

author = "Alexander Leitner and Joachimiak, {Lukasz A.} and Andreas Bracher and Leonie M{\"o}nkemeyer and Thomas Walzthoeni and Bryan Chen and Sebastian Pechmann and Susan Holmes and Yao Cong and Boxue Ma and Steve Ludtke and Wah Chiu and Hartl, {F. Ulrich} and Ruedi Aebersold and Judith Frydman",

note = "Funding Information: This work was supported by the National Institutes of Health (NIH; grants to J.F. [R01GM74074], S.J.L. [PN1EY016525], W.C. [PN1EY016525], and S.H. [R01GM086884]) and an NIH fellowship to L.A.J. (F32GM090660), as well as the European Union Seventh Framework Program PROSPECTS (Proteomics Specification in Space and Time grant HEALTH-F4-2008-201648 to R.A. and F.U.H.); the Swiss Initiative for Systems Biology; and the ERC advanced grant “Proteomics v3.0” (grant no. 233226 to R.A.). We thank Rachel Bond for help in CCTx-Cys2 experiments and Ramya Kumar for help in bTRiC purification. Stephan Nickell and Marius Boicu helped us in the EM analysis of yTRiC. Expert assistance by Stefan Pinkert in yTRiC XL-MS data analysis is gratefully acknowledged. ",

year = "2012",

month = may,

day = "9",

doi = "10.1016/j.str.2012.03.007",

language = "English (US)",

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pages = "814--825",

journal = "Structure",

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T1 - The molecular architecture of the eukaryotic chaperonin TRiC/CCT

AU - Leitner, Alexander

AU - Joachimiak, Lukasz A.

AU - Bracher, Andreas

AU - Mönkemeyer, Leonie

AU - Walzthoeni, Thomas

AU - Chen, Bryan

AU - Pechmann, Sebastian

AU - Holmes, Susan

AU - Cong, Yao

AU - Ma, Boxue

AU - Ludtke, Steve

AU - Chiu, Wah

AU - Hartl, F. Ulrich

AU - Aebersold, Ruedi

AU - Frydman, Judith

N1 - Funding Information: This work was supported by the National Institutes of Health (NIH; grants to J.F. [R01GM74074], S.J.L. [PN1EY016525], W.C. [PN1EY016525], and S.H. [R01GM086884]) and an NIH fellowship to L.A.J. (F32GM090660), as well as the European Union Seventh Framework Program PROSPECTS (Proteomics Specification in Space and Time grant HEALTH-F4-2008-201648 to R.A. and F.U.H.); the Swiss Initiative for Systems Biology; and the ERC advanced grant “Proteomics v3.0” (grant no. 233226 to R.A.). We thank Rachel Bond for help in CCTx-Cys2 experiments and Ramya Kumar for help in bTRiC purification. Stephan Nickell and Marius Boicu helped us in the EM analysis of yTRiC. Expert assistance by Stefan Pinkert in yTRiC XL-MS data analysis is gratefully acknowledged.

PY - 2012/5/9

Y1 - 2012/5/9

N2 - TRiC/CCT is a highly conserved and essential chaperonin that uses ATP cycling to facilitate folding of approximately 10% of the eukaryotic proteome. This 1 MDa hetero-oligomeric complex consists of two stacked rings of eight paralogous subunits each. Previously proposed TRiC models differ substantially in their subunit arrangements and ring register. Here, we integrate chemical crosslinking, mass spectrometry, and combinatorial modeling to reveal the definitive subunit arrangement of TRiC. In vivo disulfide mapping provided additional validation for the crosslinking-derived arrangement as the definitive TRiC topology. This subunit arrangement allowed the refinement of a structural model using existing X-ray diffraction data. The structure described here explains all available crosslink experiments, provides a rationale for previously unexplained structural features, and reveals a surprising asymmetry of charges within the chaperonin folding chamber.

AB - TRiC/CCT is a highly conserved and essential chaperonin that uses ATP cycling to facilitate folding of approximately 10% of the eukaryotic proteome. This 1 MDa hetero-oligomeric complex consists of two stacked rings of eight paralogous subunits each. Previously proposed TRiC models differ substantially in their subunit arrangements and ring register. Here, we integrate chemical crosslinking, mass spectrometry, and combinatorial modeling to reveal the definitive subunit arrangement of TRiC. In vivo disulfide mapping provided additional validation for the crosslinking-derived arrangement as the definitive TRiC topology. This subunit arrangement allowed the refinement of a structural model using existing X-ray diffraction data. The structure described here explains all available crosslink experiments, provides a rationale for previously unexplained structural features, and reveals a surprising asymmetry of charges within the chaperonin folding chamber.

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The molecular architecture of the eukaryotic chaperonin TRiC/CCT

Abstract

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