Cargando…
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...
Autores principales: | , , , , , , , |
---|---|
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 |
_version_ | 1783524828754477056 |
---|---|
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. |
format | Online Article Text |
id | pubmed-7175421 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | The Biophysical Society |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT mosbylewiss myosiniifilamentdynamicsinactinnetworksrevealedwithinterferometricscatteringmicroscopy AT hundtnikolas myosiniifilamentdynamicsinactinnetworksrevealedwithinterferometricscatteringmicroscopy AT younggavin myosiniifilamentdynamicsinactinnetworksrevealedwithinterferometricscatteringmicroscopy AT finebergadam myosiniifilamentdynamicsinactinnetworksrevealedwithinterferometricscatteringmicroscopy AT polinmarco myosiniifilamentdynamicsinactinnetworksrevealedwithinterferometricscatteringmicroscopy AT mayorsatyajit myosiniifilamentdynamicsinactinnetworksrevealedwithinterferometricscatteringmicroscopy AT kukuraphilipp myosiniifilamentdynamicsinactinnetworksrevealedwithinterferometricscatteringmicroscopy AT kosterdariusv myosiniifilamentdynamicsinactinnetworksrevealedwithinterferometricscatteringmicroscopy |