title: Fluorescence Resonance Energy Transfer (FRET)-based Detection of Profilin-VASP Interaction.
abstract: Profilins belong to a family of small G-actin binding proteins which are thought to assist in F-actin elongation at the leading edge of migrating cells through their interactions with a host of actin-binding proteins including Ena (enabled)/VASP (vasodilator stimulated phosphoprotein). Profilin's interactions with the major actin regulators have been studied almost exclusively using biochemical methods. Therefore spatiotemporal features of these protein-protein interactions have not been resolved so far. In this paper, we for the first time demonstrate the feasibility of GFP-based fluorescence resonance energy transfer (FRET) technique to detect VASP's interaction with profilin-1, a ubiquitously expressed member of profilin family of genes. Specifically, we performed acceptor photobleaching FRET in MDA-MB-231 breast cancer cells to show prominent VASP-Pfn1 interaction at the membrane ruffles near the leading edge.
authors.affiliation: Department of Bioengineering, University of Pittsburgh, 306 Center for Bioengineering, 300 Technology Drive, Pittsburgh, PA 15219, USA.
authors.forename: Dave
authors.initials: D
authors.lastname: Gau
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Ding
authors.forename: Catherine
authors.initials: C
authors.lastname: Baty
authors.forename: Partha
authors.initials: P
authors.lastname: Roy