Abstract
Nuclei are connected to the actin cytoskeleton for controlling its position in the cell and for mechanochemical signaling. Nesprin-2G is one of the major outer nuclear membrane proteins that links the nucleus to the actin cytoskeleton. In addition to its paired calponin homology (CH) domains, nesprin-2G interacts with actin filaments by binding the actin-bundling proteins FHOD1 and fascin. We describe methods to measure the interaction of nesprin-2G with actin filaments using an actin co-sedimentation assay and with its binding partner FHOD1 using a GST pull-down method.
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Acknowledgments
The methods described in this paper were supported by NIH grants R01GM099481 and R01 AR068636 to G.G.G.
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Antoku, S., Gundersen, G.G. (2018). Analysis of Nesprin-2 Interaction with Its Binding Partners and Actin. In: Gundersen, G., Worman, H. (eds) The LINC Complex. Methods in Molecular Biology, vol 1840. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8691-0_4
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DOI: https://doi.org/10.1007/978-1-4939-8691-0_4
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