Impact of In-Cell and In-Vitro Crowding on the Conformations and Dynamics of an Intrinsically Disordered Protein.
depletion interactions
fluorescence correlation spectroscopy
intrinsically disordered proteins
protein dynamics
single-molecule FRET
Journal
Angewandte Chemie (International ed. in English)
ISSN: 1521-3773
Titre abrégé: Angew Chem Int Ed Engl
Pays: Germany
ID NLM: 0370543
Informations de publication
Date de publication:
03 05 2021
03 05 2021
Historique:
revised:
08
02
2021
received:
18
12
2020
pubmed:
16
2
2021
medline:
10
8
2021
entrez:
15
2
2021
Statut:
ppublish
Résumé
The conformations and dynamics of proteins can be influenced by crowding from the large concentrations of macromolecules within cells. Intrinsically disordered proteins (IDPs) exhibit chain compaction in crowded solutions in vitro, but no such effects were observed in cultured mammalian cells. Here, to increase intracellular crowding, we reduced the cell volume by hyperosmotic stress and used an IDP as a crowding sensor for in-cell single-molecule spectroscopy. In these more crowded cells, the IDP exhibits compaction, slower chain dynamics, and much slower translational diffusion, indicating a pronounced concentration and length-scale dependence of crowding. In vitro, these effects cannot be reproduced with small but only with large polymeric crowders. The observations can be explained with polymer theory and depletion interactions and indicate that IDPs can diffuse much more efficiently through a crowded cytosol than a globular protein of similar dimensions.
Identifiants
pubmed: 33587794
doi: 10.1002/anie.202016804
doi:
Substances chimiques
Intrinsically Disordered Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
10724-10729Informations de copyright
© 2021 Wiley-VCH GmbH.
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