Studying Tau-Microtubule Interaction Using Single-Molecule TIRF Microscopy
| Autor: | Nassiba Bagdadi, Yasmina Saoudi, Isabelle Arnal, Virginie Stoppin-Mellet |
|---|---|
| Přispěvatelé: | Groupe d'imagerie neurofonctionnelle (GIN), Institut des Maladies Neurodégénératives [Bordeaux] (IMN), Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS)-Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), ANR-17-CE11-0026,MAMAs,Coordination des microtubules et de l'actine par les MAPs structurales(2017), Grenoble Institut des Neurosciences (GIN), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
Total internal reflection fluorescence microscope Chemistry Microtubule assembly [SDV]Life Sciences [q-bio] Fluorescence 03 medical and health sciences 0302 clinical medicine Microtubule Microscopy Biophysics Molecule Cytoskeleton 030217 neurology & neurosurgery ComputingMilieux_MISCELLANEOUS 030304 developmental biology |
| Zdroj: | Methods in Molecular Biology Methods in Molecular Biology, pp.77-91, 2020, ⟨10.1007/978-1-0716-0219-5_6⟩ Methods in Molecular Biology ISBN: 9781071602188 |
| Popis: | Microtubule architecture depends on a complex network of microtubule-associated proteins (MAPs) that act in concert to modulate microtubule assembly/disassembly and spatial arrangement. In vitro reconstitution of cytoskeleton dynamics coupled to single-molecule fluorescence assays has opened new perspectives to quantify the interaction of MAPs with microtubules. Here, we present a Total Internal Reflection Fluorescence (TIRF) microscopy-based assay enabling the characterization of Tau interaction with dynamic microtubules at the single-molecule level. We describe protein sample preparation in flow cells, single-molecule acquisitions by TIRF microscopy, and quantitative analysis of Tau oligomerization states and dwell time on microtubules. |
| Databáze: | OpenAIRE |
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