title: Effect of surface chemistry on tropomyosin binding to actin filaments on surfaces.
abstract: Reconstitution of actin filaments on surfaces for observation of filament-associated protein dynamics by fluorescence microscopy is currently an exciting field in biophysics. Here we examine the effects of attaching actin filaments to surfaces on the binding and dissociation kinetics of a fluorescence-labeled tropomyosin, a rod-shaped protein that forms continuous strands wrapping around the actin filament. Two attachment modalities of the actin to the surface are explored: where the actin filament is attached to the surface at multiple points along its length; and where the actin filament is attached at one end and aligned parallel to the surface by buffer flow. To facilitate analysis of actin-binding protein dynamics, we have developed a software tool for the viewing, tracing and analysis of filaments and co-localized species in noisy fluorescence timelapse images. Our analysis shows that the interaction of tropomyosin with actin filaments is similar for both attachment modalities. © 2016 Wiley Periodicals, Inc.
authors: ARC Centre of Excellence in Advanced Molecular Imaging and EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, Australia.
authors: Philip R
authors: PR
authors: Nicovich
authors: ARC Centre of Excellence in Advanced Molecular Imaging and EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, Australia.
authors: Miro
authors: M
authors: Janco
authors: School of Medical Sciences, University of New South Wales, Sydney, Australia.
authors: Tom
authors: T
authors: Sobey
authors: ARC Centre of Excellence in Advanced Molecular Imaging and EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, Australia.
authors: Mehul
authors: M
authors: Gajwani
authors: School of Medical Sciences, University of New South Wales, Sydney, Australia.
authors: Peyman
authors: P
authors: Obeidy
authors: Biomedical Imaging Facility, University of New South Wales, Sydney, Australia.
authors: Renee
authors: R
authors: Whan
authors: ARC Centre of Excellence in Advanced Molecular Imaging and EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, Australia.
authors: Katharina
authors: K
authors: Gaus
authors: School of Medical Sciences, University of New South Wales, Sydney, Australia.
authors: Peter W
authors: PW
authors: Gunning
authors: School of Mathematics and Statistics, University of New South Wales, Sydney, Australia.
authors: Adelle Cf
authors: AC
authors: Coster
authors: ARC Centre of Excellence in Advanced Molecular Imaging and EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, Australia.
authors: Till
authors: T
authors: Böcking
mesh_terms: D008841:Actin Cytoskeleton / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D003198:Computer Simulation; D008956:Models, Chemical; D012984:Software; D014335:Tropomyosin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D014335:Tropomyosin
keywords: actin filament dynamics; actin-binding proteins; actin-tropomyosin filament assembly; fluorescence microscopy; image analysis; in vitro reconstitution