Angewandte Chemie International Edition · 2014 · 25 citations · 25 references
Life SciencesBiochemistryMolecular BiologyPhotonicsProtein EngineeringStructural BiologyEnzyme FunctionFluorescence SpectroscopyOptical BiosensingPhotochemical SynthesisProtein Complex AssemblyProtein StructureGlutathione SystemProtein Complex NetworksProtein FluorescenceReal-time Processing
Spatial and temporal control over chemical and biological processes plays a key role in life and material sciences. Here we synthesized a two-photon-activatable glutathione (GSH) to trigger the interaction with glutathione S-transferase (GST) by light at superior spatiotemporal resolution. The compound shows fast and well-confined photoconversion into the bioactive GSH, which is free to interact with GST-tagged proteins. The GSH/GST interaction can be phototriggered, changing its affinity over several orders of magnitude into the nanomolar range. Multiplexed three-dimensional (3D) protein networks are simultaneously generated in situ through two-photon fs-pulsed laser-scanning excitation. The two-photon activation facilitates the three-dimensional assembly of protein structures in real time at hitherto unseen resolution in time and space, thus opening up new applications far beyond the presented examples.
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Two-Photon Laser Scanning Fluorescence Microscopy
Winfried Denk, James H. Strickler, Watt W. Webb · Science · 1990 · 9.4K citations