Methods for Determining the Cellular Functions of Vimentin Intermediate Filaments

Karen M. Ridge; Dale Shumaker; Amélie Robert; Caroline Hookway; Vladimir I. Gelfand; Paul A. Janmey; Jason Lowery; Ming Guo; David A. Weitz; Edward Kuczmarski; Robert D. Goldman

doi:10.1016/bs.mie.2015.09.036

Methods for Determining the Cellular Functions of Vimentin Intermediate Filaments

Karen M. Ridge^*, Dale Shumaker, Amélie Robert, Caroline Hookway, Vladimir I. Gelfand, Paul A. Janmey, Jason Lowery, Ming Guo, David A. Weitz, Edward Kuczmarski, Robert D. Goldman

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Chapter

30 Scopus citations

Abstract

The type III intermediate filament protein vimentin was once thought to function mainly as a static structural protein in the cytoskeleton of cells of mesenchymal origin. Now, however, vimentin is known to form a dynamic, flexible network that plays an important role in a number of signaling pathways. Here, we describe various methods that have been developed to investigate the cellular functions of the vimentin protein and intermediate filament network, including chemical disruption, photoactivation and photoconversion, biolayer interferometry, soluble bead binding assay, three-dimensional substrate experiments, collagen gel contraction, optical-tweezer active microrheology, and force spectrum microscopy. Using these techniques, the contributions of vimentin to essential cellular processes can be probed in ever further detail.

Original language	English (US)
Title of host publication	Methods in Enzymology
Publisher	Academic Press Inc
Pages	389-426
Number of pages	38
DOIs	https://doi.org/10.1016/bs.mie.2015.09.036
State	Published - 2016

Publication series

Name	Methods in Enzymology
Volume	568
ISSN (Print)	0076-6879
ISSN (Electronic)	1557-7988

Funding

Funding: R.D.G. is supported by grants from the National Institute of General Medical Sciences (P01GM09697), National Institutes of Health and the Hannah's Hope Fund. V.I.G. is supported by grants from the National Institute of General Medical Sciences (P01GM09697 and GM052111), National Institutes of Health. K.M.R. is supported by the National Heart, Lung, and Blood Institute (HL71643; HL124664), Department of Veterans Affairs (MERIT Award). P.A.J. is supported by grants from the National Institutes of health (GM096971 and EB017753). D.A.W. is supported by the NIH (PO1GM096971), the Harvard Materials Research Science and Engineering Center (DMR-0820484).

Keywords

Biolayer interferometry
Collagen gel contraction
Force spectrum microscopy
Gigaxonin
Optical tweezers
Photoactivation
Photoconversion
Three-dimensional substrate
Vimentin
Withaferin A

ASJC Scopus subject areas

Biochemistry
Molecular Biology

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10.1016/bs.mie.2015.09.036

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Ridge, K. M., Shumaker, D., Robert, A., Hookway, C., Gelfand, V. I., Janmey, P. A., Lowery, J., Guo, M., Weitz, D. A., Kuczmarski, E., & Goldman, R. D. (2016). Methods for Determining the Cellular Functions of Vimentin Intermediate Filaments. In Methods in Enzymology (pp. 389-426). (Methods in Enzymology; Vol. 568). Academic Press Inc. https://doi.org/10.1016/bs.mie.2015.09.036

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AU - Gelfand, Vladimir I.

AU - Janmey, Paul A.

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KW - Force spectrum microscopy

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Methods for Determining the Cellular Functions of Vimentin Intermediate Filaments

Abstract

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Funding

Keywords

ASJC Scopus subject areas

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