Protein structure determination in human cells by in-cell NMR and a reporter system to optimize protein delivery or transexpression.
Journal
Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179
Informations de publication
Date de publication:
02 12 2022
02 12 2022
Historique:
received:
21
07
2022
accepted:
11
11
2022
entrez:
2
12
2022
pubmed:
3
12
2022
medline:
7
12
2022
Statut:
epublish
Résumé
Most experimental methods for structural biology proceed in vitro and therefore the contribution of the intracellular environment on protein structure and dynamics is absent. Studying proteins at atomic resolution in living mammalian cells has been elusive due to the lack of methodologies. In-cell nuclear magnetic resonance spectroscopy (in-cell NMR) is an emerging technique with the power to do so. Here, we improved current methods of in-cell NMR by the development of a reporter system that allows monitoring the delivery of exogenous proteins into mammalian cells, a process that we called here "transexpression". The reporter system was used to develop an efficient protocol for in-cell NMR which enables spectral acquisition with higher quality for both disordered and folded proteins. With this method, the 3D atomic resolution structure of the model protein GB1 in human cells was determined with a backbone root-mean-square deviation (RMSD) of 1.1 Å.
Identifiants
pubmed: 36460747
doi: 10.1038/s42003-022-04251-6
pii: 10.1038/s42003-022-04251-6
pmc: PMC9718737
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1322Informations de copyright
© 2022. The Author(s).
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