Rapidly photoactivatable ATP probes for specific labeling of tropomyosin within the actomyosin protein complex.

作者: Souta Masuda , Takenori Tomohiro , Yasumaru Hatanaka

DOI: 10.1016/J.BMCL.2011.02.113

关键词: DiazirineAdenosine triphosphateBiotinylationPhotoaffinity labelingMyosin ATPaseMyosinTropomyosinChemistryBiochemistryActin

摘要: Tropomyosin-specific photoaffinity adenosine triphosphate (ATP) probes have been first developed, in which a diazirine moiety is incorporated into the γ-phosphate group as rapidly carbene-generating photophore. These clearly labeled tropomyosin presence of other actomyosin components, that is, myosin, actin, and troponins. The specific labeling was easily identified by selective trapping photo-incorporated ATP probe on Fe(3+)-immobilized metal ion affinity chromatography (IMAC) beads. characteristic nature tropomyosin-specific photocross-linking further confirmed with biotin-carrying derivative probe. data suggest actin filament assembly located close proximity to binding cavity myosin.

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