A modular DNA origami nanocompartment for engineering a cell-free, protein unfolding and degradation pathway.
Journal
Nature nanotechnology
ISSN: 1748-3395
Titre abrégé: Nat Nanotechnol
Pays: England
ID NLM: 101283273
Informations de publication
Date de publication:
29 Jul 2024
29 Jul 2024
Historique:
received:
25
09
2023
accepted:
28
05
2024
medline:
30
7
2024
pubmed:
30
7
2024
entrez:
29
7
2024
Statut:
aheadofprint
Résumé
Within the cell, chemical reactions are often confined and organized through a modular architecture. This facilitates the targeted localization of molecular species and their efficient translocation to subsequent sites. Here we present a cell-free nanoscale model that exploits compartmentalization strategies to carry out regulated protein unfolding and degradation. Our synthetic model comprises two connected DNA origami nanocompartments (each measuring 25 nm × 41 nm × 53 nm): one containing the protein unfolding machine, p97, and the other housing the protease chymotrypsin. We achieve the unidirectional immobilization of p97 within the first compartment, establishing a gateway mechanism that controls substrate recruitment, translocation and processing within the second compartment. Our data show that, whereas spatial confinement increases the rate of the individual reactions by up to tenfold, the physical connection of the compartmentalized enzymes into a chimera efficiently couples the two reactions and reduces off-target proteolysis by almost sixfold. Hence, our modular approach may serve as a blueprint for engineering artificial nanofactories with reshaped catalytic performance and functionalities beyond those observed in natural systems.
Identifiants
pubmed: 39075293
doi: 10.1038/s41565-024-01738-7
pii: 10.1038/s41565-024-01738-7
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : CRC 1096, project A6
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : 427981116
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : CRC 1430 (424228829)
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : CRC 1430 (424228829)
Informations de copyright
© 2024. The Author(s).
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