Proximity proteomics reveals a mechanism of fatty acid transfer at lipid droplet-mitochondria- endoplasmic reticulum contact sites

Abstract Membrane contact sites between organelles are critical for the transfer of biomolecules. Lipid droplets store fatty acids and form contacts with mitochondria, which regulate fatty acid oxidation and adenosine triphosphate production. Protein compartmentalization at lipid droplet-mitochondri...

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Main Authors: Ayenachew Bezawork-Geleta, Camille J. Devereux, Stacey N. Keenan, Jieqiong Lou, Ellie Cho, Shuai Nie, David P. De Souza, Vinod K. Narayana, Nicole A. Siddall, Carlos H. M. Rodrigues, Stephanie Portelli, Tenghao Zheng, Hieu T. Nim, Mirana Ramialison, Gary R. Hime, Garron T. Dodd, Elizabeth Hinde, David B. Ascher, David A. Stroud, Matthew J. Watt
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-57405-5
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author Ayenachew Bezawork-Geleta
Camille J. Devereux
Stacey N. Keenan
Jieqiong Lou
Ellie Cho
Shuai Nie
David P. De Souza
Vinod K. Narayana
Nicole A. Siddall
Carlos H. M. Rodrigues
Stephanie Portelli
Tenghao Zheng
Hieu T. Nim
Mirana Ramialison
Gary R. Hime
Garron T. Dodd
Elizabeth Hinde
David B. Ascher
David A. Stroud
Matthew J. Watt
author_facet Ayenachew Bezawork-Geleta
Camille J. Devereux
Stacey N. Keenan
Jieqiong Lou
Ellie Cho
Shuai Nie
David P. De Souza
Vinod K. Narayana
Nicole A. Siddall
Carlos H. M. Rodrigues
Stephanie Portelli
Tenghao Zheng
Hieu T. Nim
Mirana Ramialison
Gary R. Hime
Garron T. Dodd
Elizabeth Hinde
David B. Ascher
David A. Stroud
Matthew J. Watt
author_sort Ayenachew Bezawork-Geleta
collection DOAJ
description Abstract Membrane contact sites between organelles are critical for the transfer of biomolecules. Lipid droplets store fatty acids and form contacts with mitochondria, which regulate fatty acid oxidation and adenosine triphosphate production. Protein compartmentalization at lipid droplet-mitochondria contact sites and their effects on biological processes are poorly described. Using proximity-dependent biotinylation methods, we identify 71 proteins at lipid droplet-mitochondria contact sites, including a multimeric complex containing extended synaptotagmin (ESYT) 1, ESYT2, and VAMP Associated Protein B and C (VAPB). High resolution imaging confirms localization of this complex at the interface of lipid droplet-mitochondria-endoplasmic reticulum where it likely transfers fatty acids to enable β-oxidation. Deletion of ESYT1, ESYT2 or VAPB limits lipid droplet-derived fatty acid oxidation, resulting in depletion of tricarboxylic acid cycle metabolites, remodeling of the cellular lipidome, and induction of lipotoxic stress. These findings were recapitulated in Esyt1 and Esyt2 deficient mice. Our study uncovers a fundamental mechanism that is required for lipid droplet-derived fatty acid oxidation and cellular lipid homeostasis, with implications for metabolic diseases and survival.
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spelling doaj-art-3eefcf64f56544bca1b0553f1420eff52025-08-20T03:06:00ZengNature PortfolioNature Communications2041-17232025-03-0116112310.1038/s41467-025-57405-5Proximity proteomics reveals a mechanism of fatty acid transfer at lipid droplet-mitochondria- endoplasmic reticulum contact sitesAyenachew Bezawork-Geleta0Camille J. Devereux1Stacey N. Keenan2Jieqiong Lou3Ellie Cho4Shuai Nie5David P. De Souza6Vinod K. Narayana7Nicole A. Siddall8Carlos H. M. Rodrigues9Stephanie Portelli10Tenghao Zheng11Hieu T. Nim12Mirana Ramialison13Gary R. Hime14Garron T. Dodd15Elizabeth Hinde16David B. Ascher17David A. Stroud18Matthew J. Watt19Department of Anatomy and Physiology, The University of MelbourneDepartment of Anatomy and Physiology, The University of MelbourneDepartment of Anatomy and Physiology, The University of MelbourneSchool of Physics, The University of MelbourneBiological Optical Microscopy Platform (BOMP), The University of MelbourneMelbourne Mass Spectrometry and Proteomics Facility (MMSPF), Bio21 Molecular Science and Biotechnology Institute, University of MelbourneMetabolomics Australia, Bio21 Molecular Science and Biotechnology Institute, University of MelbourneMetabolomics Australia, Bio21 Molecular Science and Biotechnology Institute, University of MelbourneDepartment of Anatomy and Physiology, The University of MelbourneSchool of Chemistry and Molecular Biosciences, University of QueenslandSchool of Chemistry and Molecular Biosciences, University of QueenslandSchool of Biological Sciences, Monash UniversityMurdoch Children’s Research Institute, reNEW Novo Nordisk Foundation for Stem Cell MedicineMurdoch Children’s Research Institute, reNEW Novo Nordisk Foundation for Stem Cell MedicineDepartment of Anatomy and Physiology, The University of MelbourneDepartment of Anatomy and Physiology, The University of MelbourneSchool of Physics, The University of MelbourneSchool of Chemistry and Molecular Biosciences, University of QueenslandMurdoch Children’s Research Institute, reNEW Novo Nordisk Foundation for Stem Cell MedicineDepartment of Anatomy and Physiology, The University of MelbourneAbstract Membrane contact sites between organelles are critical for the transfer of biomolecules. Lipid droplets store fatty acids and form contacts with mitochondria, which regulate fatty acid oxidation and adenosine triphosphate production. Protein compartmentalization at lipid droplet-mitochondria contact sites and their effects on biological processes are poorly described. Using proximity-dependent biotinylation methods, we identify 71 proteins at lipid droplet-mitochondria contact sites, including a multimeric complex containing extended synaptotagmin (ESYT) 1, ESYT2, and VAMP Associated Protein B and C (VAPB). High resolution imaging confirms localization of this complex at the interface of lipid droplet-mitochondria-endoplasmic reticulum where it likely transfers fatty acids to enable β-oxidation. Deletion of ESYT1, ESYT2 or VAPB limits lipid droplet-derived fatty acid oxidation, resulting in depletion of tricarboxylic acid cycle metabolites, remodeling of the cellular lipidome, and induction of lipotoxic stress. These findings were recapitulated in Esyt1 and Esyt2 deficient mice. Our study uncovers a fundamental mechanism that is required for lipid droplet-derived fatty acid oxidation and cellular lipid homeostasis, with implications for metabolic diseases and survival.https://doi.org/10.1038/s41467-025-57405-5
spellingShingle Ayenachew Bezawork-Geleta
Camille J. Devereux
Stacey N. Keenan
Jieqiong Lou
Ellie Cho
Shuai Nie
David P. De Souza
Vinod K. Narayana
Nicole A. Siddall
Carlos H. M. Rodrigues
Stephanie Portelli
Tenghao Zheng
Hieu T. Nim
Mirana Ramialison
Gary R. Hime
Garron T. Dodd
Elizabeth Hinde
David B. Ascher
David A. Stroud
Matthew J. Watt
Proximity proteomics reveals a mechanism of fatty acid transfer at lipid droplet-mitochondria- endoplasmic reticulum contact sites
Nature Communications
title Proximity proteomics reveals a mechanism of fatty acid transfer at lipid droplet-mitochondria- endoplasmic reticulum contact sites
title_full Proximity proteomics reveals a mechanism of fatty acid transfer at lipid droplet-mitochondria- endoplasmic reticulum contact sites
title_fullStr Proximity proteomics reveals a mechanism of fatty acid transfer at lipid droplet-mitochondria- endoplasmic reticulum contact sites
title_full_unstemmed Proximity proteomics reveals a mechanism of fatty acid transfer at lipid droplet-mitochondria- endoplasmic reticulum contact sites
title_short Proximity proteomics reveals a mechanism of fatty acid transfer at lipid droplet-mitochondria- endoplasmic reticulum contact sites
title_sort proximity proteomics reveals a mechanism of fatty acid transfer at lipid droplet mitochondria endoplasmic reticulum contact sites
url https://doi.org/10.1038/s41467-025-57405-5
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