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PreprintCited by 3Mitochondrial Function and PathologyATP Synthase and ATPases ResearchAdipose Tissue and Metabolism

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A Novel Model for Proton Transport Mediated by Uncoupling Protein 1

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Jacobsen, Luise, Menon, Sneha, Gaudry, Michael J, Hakami Zanjani, Ali Asghar, Reinholdt, Peter, Jastroch, Martin, Khandelia, Himanshu·2025·Protein Science
DOI 10.1101/2025.04.28.650983

Preprint — published as 10.1002/pro.70357 · Paper page

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Abstract

Uncoupling Protein 1 (UCP1) is a mitochondrial protein which drives thermogen-esis in brown adipose tissue. UCP1 facilitates the dissipation of the proton gradient as heat and plays a critical role in energy expenditure and metabolic regulation. We employ advanced molecular simulations and mutagenesis to reveal the mechanism of UCP1-mediated proton and fatty acid (FA) transport. We demonstrate that FAs bind spontaneously to UCP1’s central substrate-binding site. In the binding site, a proton transfer to the FA is facilitated by a key aspartate residue (D28) and a coordinating water molecule. The protonated FA exits UCP1 through a well defined pathway, and releases its proton into the mitochondrial matrix. UCP1 then facilitates the return of deprotonated FAs to the intermembrane space. Nucleotide binding disrupts this mechanism by inducing conformational changes in the transmembrane helices and ob-structing the FA return pathway. Our mechanism explains every step of the transport cycle, is supported by simulation and biochemical data, and explains a diverse set of biochemical data about the transport mechanisms in UCP1 and its analogues: ANT, UCP2 and UCP3.

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Metadata

Authors
Jacobsen, L., Menon, S., Gaudry, M. J., Hakami Zanjani, A. A., Reinholdt, P., Jastroch, M., & Khandelia, H.
Affiliations
  • University of Southern Denmark (DK)
  • Stockholm University (SE)
  • Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, 5230 Odense M, Denmark
Corresponding author
Khandelia, Himanshu
ORCID
Venue
Protein Science
Publisher
openRxiv
Published
2025-05-01
Volume
34
Issue
11
Pages
e70357-e70357
ISSN
0961-8368 (ISSN-L)
DOI
10.1101/2025.04.28.650983
OpenAlex
W4415665472
Cited by
3
References
1
Citations by year
2026: 2 · 2025: 1
FWCI
0.90
Type
journal article
Language
en
Open access
Licence
Locations
  • Protein Science (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • University of Southern Denmark Research Portal (University of Southern Denmark) (submittedVersion, cc-by-nc-nd)
Funders
  • Lundbeckfonden (R344‐2020‐1023)
  • Vetenskapsrådet (2022-03136, 2024-06689)
  • Danmarks Frie Forskningsfond (3103‐00079B)
  • Danish e-Infrastructure Cooperation (DeiC-SDU-S5-202400009, DeiC-AU-N5-2023014)
  • Novo Nordisk Fonden (NNF18OC0032608, NNF20OC0065368, 0059646)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
  • Health Sciences › Medicine › Physiology (medical) › Adipose Tissue and Metabolism
Keywords
UCP1, proton transport, fatty acid uptake, brown adipose tissue thermogenesis, mitochondrial membrane transport, substrate binding site, proton transfer, aspartic acid residues, nucleotide binding, conformational change, mitochondrial matrix, energy expenditure, UCP2, UCP3, antiporters, molecular simulation
MeSH terms
Uncoupling Protein 1, Animals, Binding Sites, Fatty Acids, Humans, Ion Channels, Models, Molecular, Protons, Mitochondrial Proteins, Molecular Dynamics Simulation

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