Vesicle capture by membrane-bound Munc13-1 requires selfassembly into discrete clusters
| Autor: | Ramalingam Venkat Kalyana Sundaram, Jeff Coleman, Shyam S. Krishnakumar, Feng Li, Sathish Ramakrishnan, Alberto T. Gatta, James E. Rothman, Frederic Pincet |
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| Přispěvatelé: | Centre de recherche sur l'hétéroepitaxie et ses applications (CRHEA), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Yale University School of Medicine, Department of Cell Biology [New Haven], Yale University School of Medicine-Howard Hughes Medical Institute (HHMI), Laboratoire Charles Coulomb (L2C), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de physique de l'ENS - ENS Paris (LPENS (UMR_8023)), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Laboratoire de physique de l'ENS - ENS Paris (LPENS), Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)-Sorbonne Université (SU)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL) |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Lipid Bilayers
membrane fusion Biophysics Nerve Tissue Proteins Biochemistry Synaptic vesicle synaptic vesicle 03 medical and health sciences 0302 clinical medicine Protein Domains Structural Biology Synaptic vesicle docking Genetics Humans Active zone neurotransmission Lipid bilayer cluster Molecular Biology 030304 developmental biology [PHYS]Physics [physics] 0303 health sciences Chemistry Vesicle Cell Membrane Lipid bilayer fusion Cell Biology Photobleaching Munc13-1 Membrane HEK293 Cells SNARE Synaptic Vesicles 030217 neurology & neurosurgery |
| Zdroj: | FEBS Letters FEBS Letters, Wiley, 2021, ⟨10.1002/1873-3468.14157⟩ |
| ISSN: | 0014-5793 1873-3468 |
| DOI: | 10.1002/1873-3468.14157⟩ |
| Popis: | International audience; Munc13-1 is a large banana-shaped soluble protein that is involved in the regulation of synaptic vesicle docking and fusion. Recent studies suggest that multiple copies of Munc13-1 form nanoassemblies in active zones of neurons. However, it is not known if such clustering of Munc13-1 is correlated with multivalent binding to synaptic vesicles or specific plasma membrane domains at docking sites in the active zone. The functional significance of putative Munc13-1 clustering is also unknown. Here we report that nano-clustering is an inherent property of Munc13-1, and is indeed required for vesicle binding to bilayers containing Munc13-1. Purified Munc13-1 protein reconstituted onto supported lipid bilayers assembled into clusters containing from 2 to ~20 copies as revealed by a combination of quantitative TIRF microscopy and step-wise photobleaching. Surprisingly, only clusters containing a minimum of 6 copies of Munc13-1 were capable of efficiently capturing and retaining small unilamellar vesicles. The C-terminal C 2 C domain of Munc13-1 is not required for Munc13-1 clustering, but is required for efficient vesicle capture. This capture is largely due to a combination of electrostatic and hydrophobic interactions between the C 2 C domain and the vesicle membrane. |
| Databáze: | OpenAIRE |
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