8P4O

CryoEM structure of a GroEL7-GroES7 cage with encapsulated ordered substrate MetK in the presence of ADP-BeFx

Summary for 8P4O

• Entry DOI: 10.2210/pdb8p4o/pdb
• Related: 8P4O
• EMDB information: 17420 17421 17422 17563 17564 17565 17566 17567 17568 17569 17570 17571 17572 17573
• Descriptor: Chaperonin GroEL, Co-chaperonin GroES, S-adenosylmethionine synthase, ... (8 entities in total)
• Functional Keywords: chaperonin, folding cage, proteostasis, heat shock, atpase, chaperone
• Biological source: Escherichia coli; More
• Total number of polymer chains: 15
• Total formula weight: 519524.88

Authors

Wagner, J.,Beck, F.,Bracher, A.,Caravajal, A.I.,Wan, W.,Bohn, S.,Koerner, R.,Baumeister, W.,Fernandez-Busnadiego, R.,Hartl, F.U. (deposition date: 2023-05-23, release date: 2024-07-03, Last modification date: 2024-09-25)

Primary citation

Wagner, J.,Carvajal, A.I.,Bracher, A.,Beck, F.,Wan, W.,Bohn, S.,Korner, R.,Baumeister, W.,Fernandez-Busnadiego, R.,Hartl, F.U.
Visualizing chaperonin function in situ by cryo-electron tomography.
Nature, 633:459-464, 2024

PubMed Abstract: Chaperonins are large barrel-shaped complexes that mediate ATP-dependent protein folding. The bacterial chaperonin GroEL forms juxtaposed rings that bind unfolded protein and the lid-shaped cofactor GroES at their apertures. In vitro analyses of the chaperonin reaction have shown that substrate protein folds, unimpaired by aggregation, while transiently encapsulated in the GroEL central cavity by GroES. To determine the functional stoichiometry of GroEL, GroES and client protein in situ, here we visualized chaperonin complexes in their natural cellular environment using cryo-electron tomography. We find that, under various growth conditions, around 55-70% of GroEL binds GroES asymmetrically on one ring, with the remainder populating symmetrical complexes. Bound substrate protein is detected on the free ring of the asymmetrical complex, defining the substrate acceptor state. In situ analysis of GroEL-GroES chambers, validated by high-resolution structures obtained in vitro, showed the presence of encapsulated substrate protein in a folded state before release into the cytosol. Based on a comprehensive quantification and conformational analysis of chaperonin complexes, we propose a GroEL-GroES reaction cycle that consists of linked asymmetrical and symmetrical subreactions mediating protein folding. Our findings illuminate the native conformational and functional chaperonin cycle directly within cells.

PubMed: 39169181
DOI: 10.1038/s41586-024-07843-w

Experimental method

ELECTRON MICROSCOPY (3.04 Å)