title: A computational tool for designing FRET protein biosensors by rigid-body sampling of their conformational space.
abstract: Biosensors relying on the fluorescence resonance energy transfer (FRET) between fluorescent proteins have been used for live-cell imaging of cellular events including Ca(2+) signaling. The efficiency of energy transfer between the donor and acceptor fluorescent proteins depends on the relative distance and orientation between them, which become altered by conformational changes of a fused sensory protein caused by a cellular event. In this way, changes in FRET efficiency of Ca(2+) biosensors can be correlated with Ca(2+) concentrations. The design of these FRET biosensors can be improved by modeling conformational changes before and after a cellular event. Hence, a computational tool called FPMOD was developed to predict FRET efficiency changes by constructing FRET biosensors and sampling their conformational space through rigid-body rotation. We showed with FPMOD that our computational modeling approach can qualitatively predict the FRET efficiencies of a range of biosensors, which had strong agreement with experimental results.
authors.affiliation: Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario M5S 3G9, Canada.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Pham
authors.forename: Jason
authors.initials: J
authors.lastname: Chiang
authors.forename: Isaac
authors.initials: I
authors.lastname: Li
authors.forename: Warren
authors.initials: W
authors.lastname: Shum
authors.forename: Kevin
authors.initials: K
authors.lastname: Truong
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D002118:Calcium / Q000032:analysis; D003198:Computer Simulation; D031541:Fluorescence Resonance Energy Transfer; D008164:Luminescent Proteins / Q000737:chemistry*; D008956:Models, Chemical; D011487:Protein Conformation
chemical_list: D008164:Luminescent Proteins; D002118:Calcium