Monitoring the binding and insertion of a single transmembrane protein by an insertase.
Algorithms
Cell Membrane
/ metabolism
Escherichia coli
/ genetics
Escherichia coli Proteins
/ chemistry
Kinetics
Membrane Transport Proteins
/ chemistry
Microscopy, Atomic Force
Microscopy, Confocal
Molecular Dynamics Simulation
Mutation
Protein Binding
Protein Conformation, alpha-Helical
Spectrometry, Fluorescence
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
06 12 2021
06 12 2021
Historique:
received:
10
06
2021
accepted:
11
11
2021
entrez:
7
12
2021
pubmed:
8
12
2021
medline:
11
1
2022
Statut:
epublish
Résumé
Cells employ highly conserved families of insertases and translocases to insert and fold proteins into membranes. How insertases insert and fold membrane proteins is not fully known. To investigate how the bacterial insertase YidC facilitates this process, we here combine single-molecule force spectroscopy and fluorescence spectroscopy approaches, and molecular dynamics simulations. We observe that within 2 ms, the cytoplasmic α-helical hairpin of YidC binds the polypeptide of the membrane protein Pf3 at high conformational variability and kinetic stability. Within 52 ms, YidC strengthens its binding to the substrate and uses the cytoplasmic α-helical hairpin domain and hydrophilic groove to transfer Pf3 to the membrane-inserted, folded state. In this inserted state, Pf3 exposes low conformational variability such as typical for transmembrane α-helical proteins. The presence of YidC homologues in all domains of life gives our mechanistic insight into insertase-mediated membrane protein binding and insertion general relevance for membrane protein biogenesis.
Identifiants
pubmed: 34873152
doi: 10.1038/s41467-021-27315-3
pii: 10.1038/s41467-021-27315-3
pmc: PMC8648943
doi:
Substances chimiques
Escherichia coli Proteins
0
Membrane Transport Proteins
0
YIDC protein, E coli
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
7082Informations de copyright
© 2021. The Author(s).
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