title: Rapidly photoactivatable ATP probes for specific labeling of tropomyosin within the actomyosin protein complex.
abstract: Tropomyosin-specific photoaffinity adenosine triphosphate (ATP) probes have been first developed, in which a diazirine moiety is incorporated into the γ-phosphate group as a rapidly carbene-generating photophore. These probes clearly labeled tropomyosin in the presence of other actomyosin components, that is, myosin, actin, and troponins. The specific labeling of tropomyosin was easily identified by selective trapping of the photo-incorporated ATP probe on Fe(3+)-immobilized metal ion affinity chromatography (IMAC) beads. The characteristic nature of tropomyosin-specific photocross-linking was further confirmed with a biotin-carrying derivative of the ATP probe. These data suggest that the tropomyosin on the actin filament assembly is located in close proximity to the ATP binding cavity of myosin.
authors.affiliation: Laboratory of Biorecognition Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan.
authors.forename: Souta
authors.initials: S
authors.lastname: Masuda
authors.forename: Takenori
authors.initials: T
authors.lastname: Tomohiro
authors.forename: Yasumaru
authors.initials: Y
authors.lastname: Hatanaka
mesh_terms: D000205:Actomyosin / Q000737:chemistry* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry*; D000345:Affinity Labels / Q000737:chemistry*; D001665:Binding Sites; D019921:Biotinylation; D002846:Chromatography, Affinity; D003198:Computer Simulation; D003978:Diazomethane / Q000737:chemistry; D008697:Methane / Q000031:analogs & derivatives / Q000737:chemistry; D055668:Photochemical Processes; D014335:Tropomyosin / Q000737:chemistry*
chemical_list: D000345:Affinity Labels; D014335:Tropomyosin; C030011:carbene; D003978:Diazomethane; D000255:Adenosine Triphosphate; D000205:Actomyosin; D008697:Methane