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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Fluorescence approaches for determining protein conformations, interactions and mechanisms at membranes.
TL;DR: Using multiple independent fluorescence techniques, one can determine structural information in real time and in intact membranes under native conditions that cannot be obtained by crystallography, electron microscopy and NMR techniques, among others.
Journal ArticleDOI
Two-photon uncaging: New prospects in neuroscience and cellular biology.
David Warther,Sylvestre Gug,Alexandre Specht,Frédéric Bolze,Jean-François Nicoud,Alexandre Mourot,Maurice Goeldner +6 more
TL;DR: A literature survey of two-photon uncaging of biomolecules is described in this article, including applications in cellular- and neurobiology.
Journal ArticleDOI
Rational design of small molecule fluorescent probes for biological applications
TL;DR: This review explores general factors affecting fluorophore excitation and emission spectra, molar absorption, Stokes shift, and quantum efficiency; provides guidelines for chemist to create novel probes and presents a survey of functional probes based on PeT, FRET, and environmental or photo-sensitivity.
Journal ArticleDOI
Chemical tags: Applications in live cell fluorescence imaging
TL;DR: This review focuses on the different strategies to achieve the attachment of fluorophores to proteins in live cells and cast light on the advantages and disadvantages of each individual method.
Journal ArticleDOI
Site-Specific Protein Transamination Using N-Methylpyridinium-4-carboxaldehyde
Leah S. Witus,Chawita Netirojjanakul,Kanwal S. Palla,Ellen M. Muehl,Chih-Hisang Weng,Anthony T. Iavarone,Matthew B. Francis,Matthew B. Francis +7 more
TL;DR: N-Methylpyridinium-4-carboxaldehyde benzenesulfonate salt (Rapoport's salt, RS) was identified as a highly effective transamination reagent when paired with glutamate-terminal peptides and proteins and demonstrated that RS can be used to modify the heavy chains of the wild-type antibody or to modify both the heavy and the light chains after N- terminal sequence mutation to add additional glutamate residues.
References
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Journal ArticleDOI
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Journal ArticleDOI
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TL;DR: New fluorescent indicators for Ca2+ that are genetically encoded without cofactors and are targetable to specific intracellular locations are constructed and dubbed ‘cameleons’.
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TL;DR: The green fluorescent protein (GFP) from the Pacific Northwest jellyfish Aequorea victoria has generated intense interest as a marker for gene expression and localization of gene products.
Journal ArticleDOI
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