title: Manipulating protein conformations by single-molecule AFM-FRET nanoscopy.
abstract: Combining atomic force microscopy and fluorescence resonance energy transfer spectroscopy (AFM-FRET), we have developed a single-molecule AFM-FRET nanoscopy approach capable of effectively pinpointing and mechanically manipulating a targeted dye-labeled single protein in a large sampling area and simultaneously monitoring the conformational changes of the targeted protein by recording single-molecule FRET time trajectories. We have further demonstrated an application of using this nanoscopy on manipulation of single-molecule protein conformation and simultaneous single-molecule FRET measurement of a Cy3-Cy5-labeled kinase enzyme, HPPK (6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase). By analyzing time-resolved FRET trajectories and correlated AFM force pulling curves of the targeted single-molecule enzyme, we are able to observe the protein conformational changes of a specific coordination by AFM mechanic force pulling.
authors.affiliation: Center for Photochemical Sciences, Department of Chemistry, Bowling Green State University, Bowling Green, Ohio 43403, United States.
authors.forename: Yufan
authors.initials: Y
authors.lastname: He
authors.forename: Maolin
authors.initials: M
authors.lastname: Lu
authors.forename: Jin
authors.initials: J
authors.lastname: Cao
authors.forename: H Peter
authors.initials: HP
authors.lastname: Lu
mesh_terms: D017854:Diphosphotransferases / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005898:Glass / Q000737:chemistry; D055595:Mechanical Phenomena; D008853:Microscopy / Q000379:methods*; D018625:Microscopy, Atomic Force / Q000379:methods*; D008958:Models, Molecular; D036103:Nanotechnology / Q000379:methods*; D011487:Protein Conformation
chemical_list: D017854:Diphosphotransferases; C019684:2-amino-4-hydroxy-6-hydroxymethyldihydropteridine pyrophosphokinase