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
New strategies for fluorescently labeling proteins in the study of amyloids.
TL;DR: In this paper, the authors describe the application of different labeling methods and novel fluorescent probe strategies to the study of amyloid proteins, both for in vitro biophysical experiments and for in vivo imaging.
Journal ArticleDOI
Genetic encoding of a highly photostable, long lifetime fluorescent amino acid for imaging in mammalian cells
Chloe M. Jones,D. Miklos Robkis,Robert J. Blizzard,Mika Munari,Yarra Venkatesh,Tiberiu S. Mihaila,Alex J. Eddins,Ryan A. Mehl,William N. Zagotta,Sharona E. Gordon,E. James Petersson +10 more
TL;DR: The genetic incorporation of Acd is reported using engineered pyrrolysine tRNA synthetase (RS) mutants that allow for efficient Acd incorporation in both E. coli and mammalian cells.
Journal ArticleDOI
Binuclear Ni(II)-DpaTyr complex as a high affinity probe for an oligo-aspartate Tag tethered to proteins.
TL;DR: A remarkably high-binding affinity was achieved between the Ni(II)-DpaTyr dimer 4-4Ni(II) and the D3x2 tag peptide (DDDNGDDD), a useful tag-probe pair previously reported by us.
Journal ArticleDOI
Biarsenical-tetracysteine motif as a fluorescent tag for detection in capillary electrophoresis
TL;DR: The feasibility of using the biarsenical-tetracysteine tag to fluorescent labeling of peptides and proteins in chemical separations for chemical cytometry experiments was shown by intracellular labeling and subsequent analysis of a recombinant protein possessing the tetrACYsteine motif expressed in living cells.
Journal ArticleDOI
A novel mutation in the connexin 29 gene may contribute to nonsyndromic hearing loss.
TL;DR: It is suggested that Cx29E269D has a dominant negative effect on the formation and function of the gap junction, which provides a novel molecular explanation for the role Cx 29 plays in the development of hearing loss.
References
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Journal ArticleDOI
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Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin
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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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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.
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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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