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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Soccio RE, Breslow JL (2004) Intracellular cholesterol transport. Arterioscler Thromb Vasc Biol 24(7):1150–1160
Roseman MA, Thompson TE (1980) Mechanism of the spontaneous transfer of phospholipids between bilayers. Biochemistry 19(3):439–444
Storch J, Kleinfeld AM (1986) Transfer of long-chain fluorescent free fatty acids between unilamellar vesicles. Biochemistry 25(7):1717–1726
Cheruku S, Xu Z, Dutia R et al (2006) Mechanisms of cholesterol transfer from the Niemann-Pick Type C2 protein to model membranes supports a role in lysosomal cholesterol transport. J Biol Chem 281(42):31594–31604
Xu Z, Farver W, Kodukula S, Storch J (2008) Regulation of sterol transport between membranes and NPC2. Biochemistry 47(42):11134–11143
Kim HK, Stoch J (1992) Free fatty acid transfer from rat liver fatty acid-binding protein to phospholipid vesicles. J Biol Chem 267(1):77–82
Gomori G (1942) A modification of colorimetric phosphorus determination for use with photoelectric colorimeter. J Lab Clin Med 27:955
Backer JM, Dawidowicz EA (1981) Mechanism of cholesterol exchange between phospholipid vesicles. Biochemistry 20:3805–3810
Neufeld EB, Cooney AM, Pitha J et al (1996) Intracellular trafficking of cholesterol monitored with a cyclodextrin. J Biol Chem 271:21604–21613
Liou HL, Dixit SS, Xu S et al (2006) NPC2, the protein deficient in Niemann–Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols. J Biol Chem 281(48):36710–36723
Nichols JW, Pagano RE (1981) Kinetics of soluble lipid monomer diffusion between vesicles. Biochemistry 20(10):2783–2789
Storch J, Bass NM (1990) Transfer of fluorescent fatty acids from liver and heart fatty acid-binding proteins to model membranes. J Biol Chem 265:7827–7831
Herr FM, Li E, Weinberg RB et al (1999) Differential mechanisms of retinoid transfer from cellular retinol binding proteins types I and II to phospholipid membranes. J Biol Chem 274(14):9556–9563
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).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6875-6_9
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6873-2
Online ISBN: 978-1-4939-6875-6
eBook Packages: Springer Protocols