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Title | Structural basis of substrate progression through the bacterial chaperonin cycle. |
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Journal, issue, pages | Proc Natl Acad Sci U S A, Vol. 120, Issue 50, Page e2308933120, Year 2023 |
Publish date | Dec 12, 2023 |
Authors | Scott Gardner / Michele C Darrow / Natalya Lukoyanova / Konstantinos Thalassinos / Helen R Saibil / |
PubMed Abstract | The bacterial chaperonin GroEL-GroES promotes protein folding through ATP-regulated cycles of substrate protein binding, encapsulation, and release. Here, we have used cryoEM to determine structures ...The bacterial chaperonin GroEL-GroES promotes protein folding through ATP-regulated cycles of substrate protein binding, encapsulation, and release. Here, we have used cryoEM to determine structures of GroEL, GroEL-ADP·BeF, and GroEL-ADP·AlF-GroES all complexed with the model substrate Rubisco. Our structures provide a series of snapshots that show how the conformation and interactions of non-native Rubisco change as it proceeds through the GroEL-GroES reaction cycle. We observe specific charged and hydrophobic GroEL residues forming strong initial contacts with non-native Rubisco. Binding of ATP or ADP·BeF to GroEL-Rubisco results in the formation of an intermediate GroEL complex displaying striking asymmetry in the ATP/ADP·BeF-bound ring. In this ring, four GroEL subunits bind Rubisco and the other three are in the GroES-accepting conformation, suggesting how GroEL can recruit GroES without releasing bound substrate. Our cryoEM structures of stalled GroEL-ADP·AlF-Rubisco-GroES complexes show Rubisco folding intermediates interacting with GroEL-GroES via different sets of residues. |
External links | Proc Natl Acad Sci U S A / PubMed:38064510 / PubMed Central |
Methods | EM (single particle) |
Resolution | 3.4 - 4.4 Å |
Structure data | EMDB-15939, PDB-8ba7: EMDB-15940, PDB-8ba8: EMDB-15941: CryoEM reconstruction of GroEL-ATP-Rubisco. EMDB-15942, PDB-8ba9: EMDB-15943: CryoEM reconstruction of GroEL-GroES-ADP.AlF3-Rubisco, class I. EMDB-15945: CryoEM reconstruction of GroEL-GroES-ADP.AlF3-Rubisco, class III. EMDB-15946: CryoEM reconstruction of GroEL-GroES-ADP.AlF3-Rubisco, class IV. |
Chemicals | ChemComp-BEF: ChemComp-ADP: ChemComp-MG: ChemComp-K: ChemComp-HOH: ChemComp-AF3: |
Source |
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Keywords | CHAPERONE / GroEL / Rubisco / complex / nucleotide / Chameleon / GroES |