Slow Escape from a Helical Misfolded State of the Pore-Forming Toxin Cytolysin A.


Journal

JACS Au
ISSN: 2691-3704
Titre abrégé: JACS Au
Pays: United States
ID NLM: 101775714

Informations de publication

Date de publication:
23 Aug 2021
Historique:
received: 20 04 2021
entrez: 1 9 2021
pubmed: 2 9 2021
medline: 2 9 2021
Statut: epublish

Résumé

The pore-forming toxin cytolysin A (ClyA) is expressed as a large α-helical monomer that, upon interaction with membranes, undergoes a major conformational rearrangement into the protomer conformation, which then assembles into a cytolytic pore. Here, we investigate the folding kinetics of the ClyA monomer with single-molecule Förster resonance energy transfer spectroscopy in combination with microfluidic mixing, stopped-flow circular dichroism experiments, and molecular simulations. The complex folding process occurs over a broad range of time scales, from hundreds of nanoseconds to minutes. The very slow formation of the native state occurs from a rapidly formed and highly collapsed intermediate with large helical content and nonnative topology. Molecular dynamics simulations suggest pronounced non-native interactions as the origin of the slow escape from this deep trap in the free-energy surface, and a variational enhanced path-sampling approach enables a glimpse of the folding process that is supported by the experimental data.

Identifiants

pubmed: 34467360
doi: 10.1021/jacsau.1c00175
pmc: PMC8397351
doi:

Types de publication

Journal Article

Langues

eng

Pagination

1217-1230

Informations de copyright

© 2021 The Authors. Published by American Chemical Society.

Déclaration de conflit d'intérêts

The authors declare no competing financial interest.

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Auteurs

Fabian Dingfelder (F)

Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.

Iuri Macocco (I)

Department of Physics, Trento University, Via Sommarive 14, 38123 Povo (Trento), Italy.
SISSA, Via Bonomea 265, 34136 Trieste, Italy.

Stephan Benke (S)

Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.

Daniel Nettels (D)

Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.

Pietro Faccioli (P)

Department of Physics, Trento University, Via Sommarive 14, 38123 Povo (Trento), Italy.
INFN-TIFPA, Via Sommarive 14, 38123 Povo (Trento), Italy.

Benjamin Schuler (B)

Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
Department of Physics, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.

Classifications MeSH