Correlation of membrane protein conformational and functional dynamics.
Bacterial Outer Membrane Proteins
/ chemistry
Electrophysiology
/ methods
Escherichia coli
/ chemistry
Escherichia coli Proteins
/ chemistry
Hydrogen-Ion Concentration
Ion Channel Gating
Ion Channels
/ metabolism
Lipid Bilayers
/ chemistry
Microscopy, Atomic Force
Molecular Dynamics Simulation
Porins
/ chemistry
Protein Conformation
Protein Conformation, beta-Strand
Recombinant Proteins
Spectrum Analysis
Structure-Activity Relationship
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
16 07 2021
16 07 2021
Historique:
received:
08
12
2020
accepted:
28
06
2021
entrez:
17
7
2021
pubmed:
18
7
2021
medline:
6
8
2021
Statut:
epublish
Résumé
Conformational changes in ion channels lead to gating of an ion-conductive pore. Ion flux has been measured with high temporal resolution by single-channel electrophysiology for decades. However, correlation between functional and conformational dynamics remained difficult, lacking experimental techniques to monitor sub-millisecond conformational changes. Here, we use the outer membrane protein G (OmpG) as a model system where loop-6 opens and closes the β-barrel pore like a lid in a pH-dependent manner. Functionally, single-channel electrophysiology shows that while closed states are favored at acidic pH and open states are favored at physiological pH, both states coexist and rapidly interchange in all conditions. Using HS-AFM height spectroscopy (HS-AFM-HS), we monitor sub-millisecond loop-6 conformational dynamics, and compare them to the functional dynamics from single-channel recordings, while MD simulations provide atomistic details and energy landscapes of the pH-dependent loop-6 fluctuations. HS-AFM-HS offers new opportunities to analyze conformational dynamics at timescales of domain and loop fluctuations.
Identifiants
pubmed: 34272395
doi: 10.1038/s41467-021-24660-1
pii: 10.1038/s41467-021-24660-1
pmc: PMC8285522
doi:
Substances chimiques
Bacterial Outer Membrane Proteins
0
Escherichia coli Proteins
0
Ion Channels
0
Lipid Bilayers
0
OmpG protein, E coli
0
Porins
0
Recombinant Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
4363Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM124451
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS110790
Pays : United States
Informations de copyright
© 2021. The Author(s).
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