title: Protein Micropatterning Assay: Quantitative Analysis of Protein-Protein Interactions.
abstract: Characterization, especially quantification, of protein interactions in live cells is usually not an easy endeavor. Here, we describe a straightforward method to identify and quantify the interaction of a membrane protein ("bait") and a fluorescently labeled interaction partner ("prey") (membrane-bound or cytosolic) in live cells using Total Internal Reflection Fluorescence microscopy. The bait protein is immobilized within patterns in the plasma membrane (e.g., via an antibody); the bait-prey interaction strength can be quantified by determining the prey bulk fluorescence intensity with respect to the bait patterns. This method is particularly suitable also for the analysis of weak, transient interactions that are not easily accessible with other methods.
authors.affiliation: Institute of Applied Physics, TU Wien, Getreidemarkt 9, 1060, Vienna, Austria. schuetz@iap.tuwien.ac.at.
authors.forename: Gerhard J
authors.initials: GJ
authors.lastname: Schütz
authors.affiliation: School of Engineering and Environmental Sciences, University of Applied Sciences Upper Austria, Stelzhamerstrasse 23, 4600, Wels, Austria.
authors.forename: Julian
authors.initials: J
authors.lastname: Weghuber
authors.affiliation: School of Engineering and Environmental Sciences, University of Applied Sciences Upper Austria, Stelzhamerstrasse 23, 4600, Wels, Austria.
authors.forename: Peter
authors.initials: P
authors.lastname: Lanzerstorfer
authors.affiliation: Institute of Applied Physics, TU Wien, Getreidemarkt 9, 1060, Vienna, Austria.
authors.forename: Eva
authors.initials: E
authors.lastname: Sevcsik
mesh_terms: D005455:Fluorescent Antibody Technique; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000378:metabolism*; D057188:Workflow
chemical_list: D008565:Membrane Proteins; D011506:Proteins
keywords: Membrane proteins; Micropatterning; Protein–protein interactions; Quantitive analysis; Soft lithography; TIRF microscopy