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Myosin II Filament Dynamics in Actin Networks Revealed with Interferometric Scattering Microscopy

The plasma membrane and the underlying cytoskeletal cortex constitute active platforms for a variety of cellular processes. Recent work has shown that the remodeling acto-myosin network modifies local membrane organization, but the molecular details are only partly understood because of difficulties...

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Autores principales: Mosby, Lewis S., Hundt, Nikolas, Young, Gavin, Fineberg, Adam, Polin, Marco, Mayor, Satyajit, Kukura, Philipp, Köster, Darius V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Biophysical Society 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7175421/
https://www.ncbi.nlm.nih.gov/pubmed/32191863
http://dx.doi.org/10.1016/j.bpj.2020.02.025
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author Mosby, Lewis S.
Hundt, Nikolas
Young, Gavin
Fineberg, Adam
Polin, Marco
Mayor, Satyajit
Kukura, Philipp
Köster, Darius V.
author_facet Mosby, Lewis S.
Hundt, Nikolas
Young, Gavin
Fineberg, Adam
Polin, Marco
Mayor, Satyajit
Kukura, Philipp
Köster, Darius V.
author_sort Mosby, Lewis S.
collection PubMed
description The plasma membrane and the underlying cytoskeletal cortex constitute active platforms for a variety of cellular processes. Recent work has shown that the remodeling acto-myosin network modifies local membrane organization, but the molecular details are only partly understood because of difficulties with experimentally accessing the relevant time and length scales. Here, we use interferometric scattering microscopy to investigate a minimal acto-myosin network linked to a supported lipid bilayer membrane. Using the magnitude of the interferometric contrast, which is proportional to molecular mass, and fast acquisition rates, we detect and image individual membrane-attached actin filaments diffusing within the acto-myosin network and follow individual myosin II filament dynamics. We quantify myosin II filament dwell times and processivity as functions of ATP concentration, providing experimental evidence for the predicted ensemble behavior of myosin head domains. Our results show how decreasing ATP concentrations lead to both increasing dwell times of individual myosin II filaments and a global change from a remodeling to a contractile state of the acto-myosin network.
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spelling pubmed-71754212020-10-10 Myosin II Filament Dynamics in Actin Networks Revealed with Interferometric Scattering Microscopy Mosby, Lewis S. Hundt, Nikolas Young, Gavin Fineberg, Adam Polin, Marco Mayor, Satyajit Kukura, Philipp Köster, Darius V. Biophys J Article The plasma membrane and the underlying cytoskeletal cortex constitute active platforms for a variety of cellular processes. Recent work has shown that the remodeling acto-myosin network modifies local membrane organization, but the molecular details are only partly understood because of difficulties with experimentally accessing the relevant time and length scales. Here, we use interferometric scattering microscopy to investigate a minimal acto-myosin network linked to a supported lipid bilayer membrane. Using the magnitude of the interferometric contrast, which is proportional to molecular mass, and fast acquisition rates, we detect and image individual membrane-attached actin filaments diffusing within the acto-myosin network and follow individual myosin II filament dynamics. We quantify myosin II filament dwell times and processivity as functions of ATP concentration, providing experimental evidence for the predicted ensemble behavior of myosin head domains. Our results show how decreasing ATP concentrations lead to both increasing dwell times of individual myosin II filaments and a global change from a remodeling to a contractile state of the acto-myosin network. The Biophysical Society 2020-04-21 2020-03-04 /pmc/articles/PMC7175421/ /pubmed/32191863 http://dx.doi.org/10.1016/j.bpj.2020.02.025 Text en © 2020 Biophysical Society. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Mosby, Lewis S.
Hundt, Nikolas
Young, Gavin
Fineberg, Adam
Polin, Marco
Mayor, Satyajit
Kukura, Philipp
Köster, Darius V.
Myosin II Filament Dynamics in Actin Networks Revealed with Interferometric Scattering Microscopy
title Myosin II Filament Dynamics in Actin Networks Revealed with Interferometric Scattering Microscopy
title_full Myosin II Filament Dynamics in Actin Networks Revealed with Interferometric Scattering Microscopy
title_fullStr Myosin II Filament Dynamics in Actin Networks Revealed with Interferometric Scattering Microscopy
title_full_unstemmed Myosin II Filament Dynamics in Actin Networks Revealed with Interferometric Scattering Microscopy
title_short Myosin II Filament Dynamics in Actin Networks Revealed with Interferometric Scattering Microscopy
title_sort myosin ii filament dynamics in actin networks revealed with interferometric scattering microscopy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7175421/
https://www.ncbi.nlm.nih.gov/pubmed/32191863
http://dx.doi.org/10.1016/j.bpj.2020.02.025
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