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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A Double-Clicking Bis-Azide Fluorogenic Dye for Bioorthogonal Self-Labeling Peptide Tags
Orsolya Demeter,Eszter A. Fodor,Mihály Kállay,Gábor Mező,Krisztina Németh,Pál Szabó,Péter Kele +6 more
TL;DR: This proof-of-principle study reports on the synthesis and fluorogenic characterization of a new, double-quenched, bis-azide fluorogenic probe suitable for bioorthogonal two-point tagging of small peptide tags by double strain-promoted azide-alkyne cycloaddition.
Journal ArticleDOI
Proteomics in living cells
TL;DR: This review describes promising experimental strategies that have been devised to visualize the dynamic nature of proteins and protein complex formation occurring in living cells.
Journal ArticleDOI
Tracking the green invaders: advances in imaging virus infection in plants
Jens Tilsner,Karl Oparka +1 more
TL;DR: Technical advances that enable imaging of the infection process at previously unobtainable levels are described and the advantages and disadvantages of such techniques are described to place them in the broader perspective of their utility in analysing plant virus infection.
Journal ArticleDOI
A novel missense mutation in the connexin30 causes nonsyndromic hearing loss.
TL;DR: Findings suggest that the p.A40V mutation in CX30 causes autosomal-dominant nonsyndromic hearing loss and provides a novel molecular explanation for the role of GJB6 in hearing loss.
Journal ArticleDOI
Fluorescent labeling of proteins in living cells using the FKBP12 (F36V) tag.
Matthew B. Robers,Patrick Pinson,Louis Leong,Robert H. Batchelor,Kyle R. Gee,Thomas Machleidt +5 more
TL;DR: The optimization of a live cell labeling technology based on the use of a mutant form of FKBP12 (FKBP12(F36V) in combination with a synthetic high affinity ligand (SLF') that specifically binds to this mutant is shown.
References
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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.
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Roger Heim,Roger Y. Tsien +1 more
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