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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Live-cell protein labelling with nanometre precision by cell squeezing.
Alina Kollmannsperger,Armon Sharei,Anika Raulf,Mike Heilemann,Robert Langer,Klavs F. Jensen,Ralph Wieneke,Robert Tampé +7 more
TL;DR: In this paper, the authors describe high-throughput protein labeling facilitated by minimalistic probes delivered to mammalian cells by microfluidic cell squeezing, and demonstrate high affinity and target-specific tracing of proteins in various subcellular compartments.
Journal ArticleDOI
Manipulating proteins with chemistry: a cross-section of chemical biology
Michael E. Hahn,Tom W. Muir +1 more
TL;DR: With the development of these chemical biology tools, a time when detailed quantitative analysis of protein function, to a degree previously available only in reconstituted systems, is attainable in an in vivo setting is approaching.
Journal ArticleDOI
Myosin Va and microtubule-based motors are required for fast axonal retrograde transport of tetanus toxin in motor neurons
TL;DR: It is proposed that coordination of myosin Va and microtubule-dependent motors is required for fast axonal retrograde transport in motor neurons.
Journal ArticleDOI
Resolution of de novo HIV production and trafficking in immature dendritic cells.
Stuart Turville,Stuart Turville,Meropi Aravantinou,Hella Stössel,Nikolaus Romani,Melissa Robbiani +5 more
TL;DR: A replication-competent virus capable of being tracked preferentially within infected leukocytes is characterized and the dynamic nature of the HIV production and transfer in primary DCs is observed.
Journal ArticleDOI
Chemistry Is Dead. Long Live Chemistry
TL;DR: The recent advances in biochemistry, protein engineering, and organic synthesis that have allowed a triumphant return of chemical fluorophores to modern biological imaging are reviewed.
References
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Journal ArticleDOI
The green fluorescent protein
TL;DR: In just three years, the green fluorescent protein from the jellyfish Aequorea victoria has vaulted from obscurity to become one of the most widely studied and exploited proteins in biochemistry and cell biology.
Journal ArticleDOI
Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability.
François Denizot,Rita Lang +1 more
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Journal ArticleDOI
Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin
Atsushi Miyawaki,Juan Llopis,Roger Heim,J. Michael McCaffery,Joseph A. Adams,Mitsuhiko Ikura,Mitsuhiko Ikura,Roger Y. Tsien +7 more
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’.
Journal ArticleDOI
Crystal structure of the Aequorea victoria green fluorescent protein.
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
Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer
Roger Heim,Roger Y. Tsien +1 more
TL;DR: The results demonstrate that the production of more and better GFP variants is possible and worthwhile, and facilitates multicolor imaging of differential gene expression, protein localization or cell fate.
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