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Sustained enzymatic activity and flow in crowded protein droplets

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Testa, Andrea, Dindo, Mirco, Rebane, Aleksander A., Nasouri, Babak, Style, Robert W., Golestanian, Ramin, Dufresne, Eric R., Laurino, Paola·2021·Nature Communications
DOI 10.1038/s41467-021-26532-0PMC PMC8560906

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Abstract

Living cells harvest energy from their environments to drive the chemical processes that enable life. We introduce a minimal system that operates at similar protein concentrations, metabolic densities, and length scales as living cells. This approach takes advantage of the tendency of phase-separated protein droplets to strongly partition enzymes, while presenting minimal barriers to transport of small molecules across their interface. By dispersing these microreactors in a reservoir of substrate-loaded buffer, we achieve steady states at metabolic densities that match those of the hungriest microorganisms. We further demonstrate the formation of steady pH gradients, capable of driving microscopic flows. Our approach enables the investigation of the function of diverse enzymes in environments that mimic cytoplasm, and provides a flexible platform for studying the collective behavior of matter driven far from equilibrium.

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Metadata

Authors
Testa, A., Dindo, M., Rebane, A. A., Nasouri, B., Style, R. W., Golestanian, R., Dufresne, E. R., & Laurino, P.
Affiliations
  • ETH Zurich (CH)
  • Okinawa Institute of Science and Technology Graduate University (JP)
  • Max Planck Institute for Dynamics and Self-Organization (DE)
  • University of Oxford (GB)
Corresponding author
  • Dufresne, Eric R.
  • Laurino, Paola
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2021-11-01
Volume
12
Issue
1
Pages
6293-6293
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-021-26532-0
PMC
PMC8560906
OpenAlex
W3210793832
Cited by
144
References
10
Citations by year
2026: 27 · 2025: 39 · 2024: 42 · 2023: 25 · 2022: 11
FWCI
10.48
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Institutional Repositories DataBase (IRDB) (submittedVersion, cc-by-nc-nd)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Okinawa Institute of Science and Technology Graduate University (Okinawa Institute of Science and Technology Graduate University) (submittedVersion, cc-by-nc-nd)
  • Repository for Publications and Research Data (ETH Zurich) (submittedVersion, cc-by)
  • ETH Zürich Research Collection (cc-by)
Funders
Okinawa Institute of Science and Technology Graduate University
Topics
  • Physical Sciences › Physics and Astronomy › Astronomy and Astrophysics › Origins and Evolution of Life
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Physical Sciences › Chemical Engineering › Fluid Flow and Transfer Processes › Innovative Microfluidic and Catalytic Techniques Innovation
Keywords
protein droplets, phase separation, enzyme activity, enzyme partitioning, substrate transport, pH gradient, microfluidic flow, far-from-equilibrium systems
MeSH terms
Catalysis, Humans, Hydro-Lyases, Models, Biological, Urease, Nanoparticles, Artificial Cells

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