Journal ArticleDOI
Specific Covalent Labeling of Recombinant Protein Molecules Inside Live Cells
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TLDR
This system provides a recipe for slightly modifying a target protein so that it can be singled out from the many other proteins inside live cells and fluorescently stained by small nonfluorescent dye molecules added from outside the cells.Abstract:
Recombinant proteins containing four cysteines at the i , i + 1, i + 4, and i + 5 positions of an α helix were fluorescently labeled in living cells by extracellular administration of 4′,5′-bis(1,3,2-dithioarsolan-2-yl)fluorescein. This designed small ligand is membrane-permeant and nonfluorescent until it binds with high affinity and specificity to the tetracysteine domain. Such in situ labeling adds much less mass than does green fluorescent protein and offers greater versatility in attachment sites as well as potential spectroscopic and chemical properties. This system provides a recipe for slightly modifying a target protein so that it can be singled out from the many other proteins inside live cells and fluorescently stained by small nonfluorescent dye molecules added from outside the cells.read more
Citations
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Journal ArticleDOI
Fluorescent labeling of tetracysteine-tagged proteins in intact cells
Carsten Hoffmann,Guido M. Gaietta,Alexander Zürn,Stephen R. Adams,Sonia Terrillon,Mark H. Ellisman,Roger Y. Tsien,Martin J. Lohse +7 more
TL;DR: A generally applicable labeling procedure that can be applied to proteins with expression below 1 pmol mg−1 of protein, such as G protein–coupled receptors, and it can be used to study the intracellular localization of proteins as well as functional interactions in fluorescence resonance energy transfer experiments.
Journal ArticleDOI
Chemical tags for labeling proteins inside living cells.
Chaoran Jing,Virginia W. Cornish +1 more
TL;DR: An overview of different chemical tagging strategies and technologies is presented and the challenge of rendering the labeling reaction sufficiently selective and the fluorophore probe sufficiently well behaved to image intracellular proteins with high signal-to-noise ratios is emphasized.
Journal ArticleDOI
Transgenically Encoded Protein Photoinactivation (FlAsH-FALI): Acute Inactivation of Synaptotagmin I
Kurt W. Marek,Graeme W. Davis +1 more
TL;DR: It is demonstrated that Syt I is required for a post-docking step during vesicle fusion but does not function to stabilize the docked vesicles state.
Journal ArticleDOI
Three-dimensional harmonic holographic microcopy using nanoparticles as probes for cell imaging
TL;DR: High-resolution 3D distributions of these SHG markers in mammalian cells are successfully captured and interpreted by the H(2) microscope and the coherent SHG signal from BaTiO(3) nanoparticles is used for three-dimensional (3D) imaging without scanning.
Journal ArticleDOI
Coronavirus envelope (E) protein remains at the site of assembly.
TL;DR: The results provide strong support that E proteins carry out their function(s) at the site of budding/assembly in CoV A59 and confirm the presence of E in Golgi cisternae.
References
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