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
Site-selective covalent reactions on proteinogenic amino acids
TL;DR: Recent advancements of bioconjugation reactions that demonstrate the feature of precise site selectivity in the reactions of the proteinogenic amino acids are summarized, focusing on those at non-coded or non-proteinogenic amino acid that are introduced to proteins through genetic manipulations.
Journal ArticleDOI
The switching of a Rhenium(I) complex from turn-off to turn-on sensor system through protein binding
Jayaraman Bhuvaneswari,Paulpandian Muthu Mareeswaran,Paulpandian Muthu Mareeswaran,Karunanithi Anandababu,Seenivasan Rajagopal +4 more
TL;DR: In this article, luminescent Re(I)-polypyridine complexes (1a and 1b) containing an amide pyridine pendant carrying the N-H group have been synthesized, characterized and used as an anion sensor by utilizing UV-visible and luminescence titration methods.
Journal ArticleDOI
Imaging techniques in plant transport: Meeting review
Mark D. Fricker,K J Oparka +1 more
TL;DR: UV laser-light provide an equally powerful tool either to ablate whole cells completely for developmental or to punch-out tiny holes in the cell wall to give access to the plasma membrane.
Journal ArticleDOI
T-CrAsH: a heterologous chemical crosslinker.
Anna Rutkowska,Tilman Plass,Jan Erik Hoffmann,Dmytro A. Yushchenko,Suihan Feng,Carsten Schultz +5 more
TL;DR: By combining strain‐promoted Diels–Alder cycloaddition with FlAsH‐based labeling of peptidic tetracysteine motifs, the membrane‐permeating reversible crosslinker T‐CrAsH is developed, demonstrating the feasibility of the method both in vitro and inside cells.
Journal ArticleDOI
Visualizing hepatitis B virus with biarsenical labelling in living cells.
Shuzhen Sun,Jingjun Yan,Chao Xia,Yuanyuan Lin,Xiaorui Jiang,Haojing Liu,Huan-ping Ren,Junwei Yan,Ju-Sheng Lin,Xing-Xing He +9 more
TL;DR: GFP tag, as a large inserted fragment, is not suitable for labelling Hepatitis B virus (HBV) that is a compact virion with limited internal space.
References
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Journal ArticleDOI
The green fluorescent protein
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Journal ArticleDOI
Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability.
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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’.
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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.
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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