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Transport Assays for Sterol-Binding Proteins: Stopped-Flow Fluorescence Methods for Investigating Intracellular Cholesterol Transport Mechanisms of NPC2 Protein

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Cholesterol Homeostasis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1583))

Abstract

In this chapter we describe the use of stopped flow fluorescence spectroscopy to analyze the kinetic mechanisms of protein mediated cholesterol transfer to, from, and between model membranes. These assays allow for the detection of protein–membrane interactions that may occur during cholesterol transfer by simply modifying donor or acceptor concentrations, membrane composition, or buffer properties, and analyzing resultant transfer rates.

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Acknowledgments

This work was supported by the Ara Parseghian Medical Research Foundation (J.S.), the American Heart Association (L.M. and J.S.), and the National Institute of Health (J.S.) (GM 115866).

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Correspondence to Judith Storch .

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McCauliff, L.A., Storch, J. (2017). Transport Assays for Sterol-Binding Proteins: Stopped-Flow Fluorescence Methods for Investigating Intracellular Cholesterol Transport Mechanisms of NPC2 Protein. In: Gelissen, I., Brown, A. (eds) Cholesterol Homeostasis. Methods in Molecular Biology, vol 1583. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6875-6_9

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  • DOI: https://doi.org/10.1007/978-1-4939-6875-6_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6873-2

  • Online ISBN: 978-1-4939-6875-6

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