Carrington, G., Takagi, Y., Umney, O. et al. (3 more authors) (2024) Expression, purification and in vitro analysis of myosin. In: Lavelle, C. and Le Gall, A., (eds.) Molecular Motors. Methods in Molecular Biology, 2881 . Humana New York, NY , pp. 167-192. ISBN 9781071642795
Abstract
To understand the mechanics and kinetic properties of cytoskeletal molecular motors such as myosin, typically the motor of interest needs to be expressed and purified and then analyzed using a range of in vitro-based assays. In this chapter, we describe how to express and purify myosin using the insect cell system, how to characterize the purified protein by mass photometry and negative-stain EM to assess its quality, and how to perform in vitro assays in which fluorescently labeled myosin walks along actin tracks, including a brief description of adapting these assays for MINFLUX imaging.
Metadata
Item Type: | Book Section |
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Authors/Creators: |
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Editors: |
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Copyright, Publisher and Additional Information: | This version of the chapter has been accepted for publication, after peer review (when applicable) and is subject to Springer Nature’s AM terms of use (https://www.springernature.com/gp/open-research/policies/accepted-manuscript-terms), but is not the Version of Record and does not reflect post-acceptance improvements, or any corrections. The Version of Record is available online at: https://doi.org/10.1007/978-1-0716-4280-1_9 |
Keywords: | Myosin; Actin; In vitro motility; Negative-stain electron microscopy; Mass photometry |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Cell Biology (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 03 Feb 2025 14:59 |
Last Modified: | 03 Feb 2025 14:59 |
Status: | Published |
Publisher: | Humana New York, NY |
Series Name: | Methods in Molecular Biology |
Identification Number: | 10.1007/978-1-0716-4280-1_9 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:222746 |
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Filename: Methods_final.pdf
