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
Studying protein dynamics in living cells.
TL;DR: Live cell imaging, in combination with photobleaching, energy transfer or fluorescence correlation spectroscopy are providing unprecedented insights into the movement of proteins and their interactions with cellular components.
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Functionalizing nanoparticles with biological molecules: developing chemistries that facilitate nanotechnology.
Kim E. Sapsford,W. Russ Algar,Lorenzo Berti,Kelly Boeneman Gemmill,Brendan J. Casey,Eunkeu Oh,Michael H. Stewart,Igor L. Medintz +7 more
TL;DR: Chemistries that Facilitate Nanotechnology Kim E. Sapsford,† W. Russ Algar, Lorenzo Berti, Kelly Boeneman Gemmill,‡ Brendan J. Casey,† Eunkeu Oh, Michael H. Stewart, and Igor L. Medintz .
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Light Microscopy Techniques for Live Cell Imaging
TL;DR: A brief overview of the main approaches to live cell imaging is given, with some mention of their pros and cons.
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A general method to improve fluorophores for live-cell and single-molecule microscopy
Jonathan B. Grimm,Brian P. English,Jiji Chen,Joel P Slaughter,Zhengjian Zhang,Andrey Revyakin,Ronak Patel,John J. Macklin,Davide Normanno,Robert H. Singer,Timothée Lionnet,Luke D. Lavis +11 more
TL;DR: Inspired by molecular modeling, the N,N-dimethylamino substituents in tetramethylrhodamine are replaced with four-membered azetidine rings, which doubles the quantum efficiency and improves the photon yield of the dye in applications ranging from in vitro single-molecule measurements to super-resolution imaging.
Journal ArticleDOI
Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin
Bijan Zakeri,Jacob O. Fierer,Emrah Celik,Emily Chittock,Ulrich Schwarz-Linek,Vincent T. Moy,Mark Howarth +6 more
TL;DR: The robust reaction conditions and irreversible linkage of SpyTag shed light on spontaneous isopeptide bond formation and should provide a targetable lock in cells and a stable module for new protein architectures.
References
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