Abstract
The disassembly of the clathrin lattice surrounding coated vesicles is the obligatory last step in their life cycle. It is mediated by the coordinated recruitment of auxilin and Hsc70, an ATP-driven molecular clamp. Here, we describe the preparation of reagents and the single-particle fluorescence microscopy imaging assay in which we visualize directly the Hsc70-driven uncoating of synthetic clathrin coats or clathrin-coated vesicles.
The original version of this chapter was revised. A correction to this chapter can be found at https://doi.org/10.1007/978-1-4939-8719-1_19
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Change history
24 October 2018
This book was inadvertently published with the incorrect title as Clathrin-Mediated Endoytosis: Methods and Protocols. This has now been corrected throughout the book to Clathrin-Mediated Endocytosis: Methods and Protocols.
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Acknowledgments
T.B. was supported by grants from the Australian Research Council (DP130100936, FT 1001004) and the National Health and Medical Research Council (1098870, 1100771).
The Kirchhausen laboratory was generously supported by grants from Biogen and by National Institutes of Health Grants R01 GM075252.
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Böcking, T., Upadhyayula, S., Rapoport, I., Capraro, B.R., Kirchhausen, T. (2018). Reconstitution of Clathrin Coat Disassembly for Fluorescence Microscopy and Single-Molecule Analysis. In: Swan, L. (eds) Clathrin-Mediated Endocytosis. Methods in Molecular Biology, vol 1847. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8719-1_10
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DOI: https://doi.org/10.1007/978-1-4939-8719-1_10
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