Peptide-PAINT Enables Investigation of Endogenous Talin with Molecular Scale Resolution in Cells and Tissues.
cell adhesion
peptides
protein-protein interactions
single-molecule studies
talin
Journal
Chembiochem : a European journal of chemical biology
ISSN: 1439-7633
Titre abrégé: Chembiochem
Pays: Germany
ID NLM: 100937360
Informations de publication
Date de publication:
01 10 2021
01 10 2021
Historique:
revised:
16
07
2021
received:
18
06
2021
pubmed:
22
7
2021
medline:
22
2
2022
entrez:
21
7
2021
Statut:
ppublish
Résumé
Talin is a cell adhesion molecule that is indispensable for the development and function of multicellular organisms. Despite its central role for many cell biological processes, suitable methods to investigate the nanoscale organization of talin in its native environment are missing. Here, we overcome this limitation by combining single-molecule resolved PAINT (points accumulation in nanoscale topography) imaging with the IRIS (image reconstruction by integrating exchangeable single-molecule localization) approach, enabling the quantitative analysis of genetically unmodified talin molecules in cells. We demonstrate that a previously reported peptide can be utilized to specifically label the two major talin isoforms expressed in mammalian tissues with a localization precision of <10 nm. Our experiments show that the methodology performs equally well as state-of-the-art single-molecule localization techniques, and the first applications reveal a thus far undescribed cell adhesion structure in differentiating stem cells. Furthermore, we demonstrate the applicability of this peptide-PAINT technique to mouse tissues paving the way to single-protein imaging of endogenous talin proteins under physiologically relevant conditions.
Identifiants
pubmed: 34286903
doi: 10.1002/cbic.202100301
pmc: PMC8518977
doi:
Substances chimiques
Peptides
0
Talin
0
Tln1 protein, mouse
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2872-2879Subventions
Organisme : WWU Münster
Organisme : Max Planck Institute of Biochemistry
Organisme : German Research Foundation
Organisme : Collaborative Research Consortium SFB
ID : 1348
Organisme : DFG-ANR program
ID : GR3399/6-1
Organisme : Human Frontiers Science Program
ID : RGP0024
Informations de copyright
© 2021 The Authors. ChemBioChem published by Wiley-VCH GmbH.
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