Academic Journal
Quantitative FRET FLIM imaging reveals multiple interactions between proteins of the ESYT, ORP, and VAP families at the endoplasmic reticulum membrane.
| Τίτλος: | Quantitative FRET FLIM imaging reveals multiple interactions between proteins of the ESYT, ORP, and VAP families at the endoplasmic reticulum membrane. |
|---|---|
| Συγγραφείς: | Zamiati D; Université Paris-Saclay, CNRS, Institut de Chimie Physique, UMR CNRS 8000, Orsay, France., Abdesselem M; Université Paris-Saclay, CNRS, Institut de Chimie Physique, UMR CNRS 8000, Orsay, France., Nüsse O; Université Paris-Saclay, CNRS, Institut de Chimie Physique, UMR CNRS 8000, Orsay, France. Electronic address: oliver.nusse@universite-paris-saclay.fr., Erard M; Université Paris-Saclay, CNRS, Institut de Chimie Physique, UMR CNRS 8000, Orsay, France. Electronic address: marie.erard@universite-paris-saclay.fr. |
| Πηγή: | The Journal of biological chemistry [J Biol Chem] 2026 Jul; Vol. 302 (7), pp. 113144. Date of Electronic Publication: 2026 May 13. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE |
| Imprint Name(s): | Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology |
| Ιατρικοί όροι (MeSH): | Endoplasmic Reticulum*/metabolism , Endoplasmic Reticulum*/chemistry , Receptors, Steroid*/metabolism , Receptors, Steroid*/chemistry , Membrane Proteins*/metabolism , Membrane Proteins*/chemistry , Vesicular Transport Proteins*/metabolism , Vesicular Transport Proteins*/chemistry , Fluorescence Resonance Energy Transfer*, Oxysterol Binding Proteins ; Humans ; Microscopy, Fluorescence ; Synaptotagmins |
| Περίληψη: | The endoplasmic reticulum (ER) is a highly dynamic intracellular organelle that forms close contact sites with other organelles and the plasma membrane. These membrane contact sites play essential roles in lipid exchange and calcium homeostasis. ER membrane proteins from the VAP, ORP, and ESYT families are key players in the formation and function of these contacts. Numerous interactions between these proteins are likely critical for their activity. We investigate the interactome between these protein families in live cells by analyzing two members from each family, using FRET between pairs of proteins labeled with fluorescent proteins. FRET is detected by Fluorescence Lifetime Imaging Microscopy. Our quantitative approach shows that all tested proteins clusterize and sometimes interact within their respective families and that their organization at the nanometer scale can be disrupted by specific mutations or domain deletions. In particular, we show that the coiled-coil domains of ORP5 and ORP8 are required for the formation of both homomeric and heteromeric complexes. Moreover, we demonstrate that FRET-FLIM can detect intramolecular conformational changes in response to alterations in the cellular environment, such as variations in Ca2+ concentration, as observed for ESYT1. We also identify novel interfamily organization, including clustering between VAPB and ORP8, and between ESYT2 and ORP5/8. Finally, our approach highlights the broad interaction network (interactome) of VAPA/B and shows how various potential binding partners can influence FRET efficiency. (Copyright © 2026 The Authors. Published by Elsevier Inc. All rights reserved.) |
| Competing Interests: | Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article. |
| Contributed Indexing: | Keywords: FRET FLIM microscopy; Oxysterol-binding protein-related proteins; Vesicle-associated membrane protein-associated proteins; endoplasmic reticulum; extended synaptotagmin; live-cell imaging; membrane contact sites; protein interactions |
| Substance Nomenclature: | 0 (Oxysterol Binding Proteins) 0 (Receptors, Steroid) 0 (Membrane Proteins) 0 (Vesicular Transport Proteins) 0 (ESYT1 protein, human) 0 (VAPA protein, human) 0 (ESYT2 protein, human) 134193-27-4 (Synaptotagmins) |
| Entry Date(s): | Date Created: 20260514 Date Completed: 20260720 Latest Revision: 20260726 |
| Update Code: | 20260726 |
| PubMed Central ID: | PMC13292579 |
| DOI: | 10.1016/j.jbc.2026.113144 |
| PMID: | 42134546 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 1083-351X |
|---|---|
| DOI: | 10.1016/j.jbc.2026.113144 |