@article {47675,
	title = {Determining membrane protein topologies in single cells and high-throughput screening applications.},
	journal = {Current protocols in cell biology / editorial board, Juan S. Bonifacino ... [et al.]},
	volume = {Chapter 5},
	year = {2010},
	month = {2010 Dec},
	pages = {Unit 5.7},
	abstract = {<p>Correct localization and topology are crucial for a protein{\textquoteright}s cellular function. To determine topologies of membrane proteins, a new technique, called fluorescence protease protection (FPP) assay, has recently been established. The sole requirements for FPP are the expression of fluorescent-protein fusion proteins and the selective permeabilization of the plasma membrane, permitting a wide range of cell types and organelles to be investigated. Proteins topologies in organelles like endoplasmic reticulum, Golgi apparatus, mitochondria, peroxisomes, and autophagosomes have already been determined by FPP. Here, two different step-by-step protocols of the FPP assay are provided. First, we describe the FPP assay using fluorescence microscopy for single adherent cells, and second, we outline the FPP assay for high-throughput screening applications.</p>
},
	keywords = {Animals, Cell Line, Cytological Techniques, High-Throughput Screening Assays, Humans, Luminescent Proteins, Membrane Proteins, Microscopy, Fluorescence, Peptide Hydrolases, Protein Transport, Recombinant Fusion Proteins, Single-Cell Analysis},
	issn = {1934-2616},
	doi = {10.1002/0471143030.cb0507s49},
	author = {Wunder, Christian and Lippincott-Schwartz, Jennifer and Lorenz, Holger}
}
