title: Transport Assays for Sterol-Binding Proteins: Stopped-Flow Fluorescence Methods for Investigating Intracellular Cholesterol Transport Mechanisms of NPC2 Protein.
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.
authors.affiliation: Department of Nutritional Sciences and Rutgers Center for Lipid Research, Rutgers University New Brunswick, 65 Dudley Road, New Brunswick, NJ, 08901, USA.
authors.forename: Leslie A
authors.initials: LA
authors.lastname: McCauliff
authors.affiliation: Department of Nutritional Sciences and Rutgers Center for Lipid Research, Rutgers University New Brunswick, 65 Dudley Road, New Brunswick, NJ, 08901, USA. storch@aesop.rutgers.edu.
authors.forename: Judith
authors.initials: J
authors.lastname: Storch
mesh_terms: D001693:Biological Transport, Active / Q000502:physiology; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002784:Cholesterol / Q000737:chemistry* / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008567:Membranes, Artificial; D013050:Spectrometry, Fluorescence; D033921:Vesicular Transport Proteins
chemical_list: D002352:Carrier Proteins; D006023:Glycoproteins; D008567:Membranes, Artificial; C104585:NPC2 protein, human; D033921:Vesicular Transport Proteins; D002784:Cholesterol
keywords: Cholesterol; Cholesterol trafficking; Collisional transfer; Diffusional transfer; FRET; Fluorescence; Model membranes; Small unilamellar vesicles; Stopped flow