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Circular Dichroism and Isothermal Titration Calorimetry to Study the Interaction of α-Synuclein with Membranes

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Alpha-Synuclein

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1948))

Abstract

α-Synuclein’s physiology and pathology have been linked by numerous reports to its ability to bind and remodel membranes, especially at synaptic terminals. It is therefore critical for researchers investigating the determinants of these interactions to rely on methods capable of providing an accurate and complete physicochemical snapshot of the binding events. Circular dichroism (CD) and isothermal titration calorimetry (ITC) are established techniques for the study of binding equilibria in biological systems and, especially when used in combination, allow a thorough characterization of the protein-lipid interplay.

Here we provide general guidelines and describe some common pitfalls of these experiments. This protocol describes the preparation of small unilamellar vesicles (SUVs), mimicking the curved bilayers α-synuclein normally interacts with, the CD-monitored titration of α-synuclein with SUVs, the ITC (lipid-into-protein) experiment, and the subsequent data analysis using an n independent binding site model.

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Acknowledgments

M.R. thanks Kelly L. Arnett and the Center for Macromolecular Interactions at the Harvard Medical School Department of Biological Chemistry and Molecular Pharmacology for assisting with CD and ITC measurements, revising this manuscript, and many helpful discussions; Alex E. Powers for helping with data collection and analysis; and Martina Astrid Rodda for editing and revising the manuscript.

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Authors and Affiliations

  1. Ann Romney Center for Neurologic Diseases, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, USA

    Matteo Rovere

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  1. Matteo Rovere
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Correspondence to Matteo Rovere .

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Editors and Affiliations

  1. Dementia Research Institute, University College London, London, UK

    Tim Bartels

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Rovere, M. (2019). Circular Dichroism and Isothermal Titration Calorimetry to Study the Interaction of α-Synuclein with Membranes. In: Bartels, T. (eds) Alpha-Synuclein. Methods in Molecular Biology, vol 1948. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9124-2_11

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  • DOI: https://doi.org/10.1007/978-1-4939-9124-2_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9123-5

  • Online ISBN: 978-1-4939-9124-2

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