Symmetry-free cryo-EM structures of the chaperonin TRiC along its ATPase-driven conformational cycle.
Yao Cong,Gunnar F. Schröder,Anne S. Meyer,Joanita Jakana,Boxue Ma,Matthew T. Dougherty,Michael F. Schmid,Stefanie Reissmann,Michael Levitt,Steven L Ludtke,Judith Frydman,Wah Chiu +11 more
TLDR
Five cryo‐EM structures of TRiC in apo and nucleotide‐induced states without imposing symmetry during the 3D reconstruction reveal the intra‐ and inter‐ring subunit interaction pattern changes during the ATPase cycle.Abstract:
The eukaryotic group II chaperonin TRiC/CCT is a 16-subunit complex with eight distinct but similar subunits arranged in two stacked rings. Substrate folding inside the central chamber is triggered by ATP hydrolysis. We present five cryo-EM structures of TRiC in apo and nucleotide-induced states without imposing symmetry during the 3D reconstruction. These structures reveal the intra- and inter-ring subunit interaction pattern changes during the ATPase cycle. In the apo state, the subunit arrangement in each ring is highly asymmetric, whereas all nucleotide-containing states tend to be more symmetrical. We identify and structurally characterize an one-ring closed intermediate induced by ATP hydrolysis wherein the closed TRiC ring exhibits an observable chamber expansion. This likely represents the physiological substrate folding state. Our structural results suggest mechanisms for inter-ring-negative cooperativity, intra-ring-positive cooperativity, and protein-folding chamber closure of TRiC. Intriguingly, these mechanisms are different from other group I and II chaperonins despite their similar architecture.read more
Citations
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Journal ArticleDOI
Molecular Chaperone Functions in Protein Folding and Proteostasis
TL;DR: This review focuses on recent advances in understanding the mechanisms of chaperone action in promoting and regulating protein folding and on the pathological consequences of protein misfolding and aggregation.
Journal ArticleDOI
Chaperone machines for protein folding, unfolding and disaggregation
TL;DR: The structural basis of their mechanism of action is being unravelled and typically involves massive displacements of 20–30 kDa domains over distances of 20-50 Å and rotations of up to 100°.
Journal ArticleDOI
Structural Probing of a Protein Phosphatase 2A Network by Chemical Cross-Linking and Mass Spectrometry
Franz Herzog,Abdullah Kahraman,Daniel Boehringer,Raymond H. Mak,Andreas Bracher,Thomas Walzthoeni,Alexander Leitner,Martin Beck,Franz-Ulrich Hartl,Nenad Ban,Lars Malmström,Ruedi Aebersold +11 more
TL;DR: This study establishes XL-MS as an integral part of hybrid structural biology approaches for the analysis of endogenous protein complexes by gaining distance restraints on a modular interaction network of protein complexes affinity-purified from human cells by applying an adaptedXL-MS protocol.
Journal ArticleDOI
The Molecular Architecture of the Eukaryotic Chaperonin TRiC/CCT
Alexander Leitner,Lukasz A. Joachimiak,Andreas Bracher,Leonie Mönkemeyer,Thomas Walzthoeni,Thomas Walzthoeni,Bryan Chen,Sebastian Pechmann,Susan Holmes,Yao Cong,Boxue Ma,S. Ludtke,Wah Chiu,F. Ulrich Hartl,Ruedi Aebersold,Ruedi Aebersold,Judith Frydman +16 more
TL;DR: This work integrates chemical crosslinking, mass spectrometry, and combinatorial modeling to reveal the definitive subunit arrangement of TRiC and 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.
Journal ArticleDOI
Chemical cross-linking/mass spectrometry targeting acidic residues in proteins and protein complexes
Alexander Leitner,Lukasz A. Joachimiak,Pia Unverdorben,Thomas Walzthoeni,Judith Frydman,Friedrich Förster,Ruedi Aebersold +6 more
TL;DR: A chemistry to cross-link acidic residues that generates structural information complementary to that obtained by amine-specific cross-linking, thus significantly expanding the scope of XL-MS analyses and expanding the yield of structural information that can be obtained from cross- linking studies and used in hybrid modeling approaches.
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