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
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Journal ArticleDOI
Thiol-Mediated Uptake
Quentin Laurent,Rémi Martinent,Bumhee Lim,Anh-Tuan Pham,Takehiro Kato,Javier López-Andarias,Naomi Sakai,Stefan Matile +7 more
TL;DR: This Perspective focuses on thiol-mediated uptake, that is, the entry of substrates into cells enabled by oligochalcogenides or mimics, often disulfides, and inhibited byThiol-reactive agents.
Journal ArticleDOI
Imaging the assembly, structure and activity of type III secretion systems.
Jost Enninga,Ilan Rosenshine +1 more
TL;DR: Light and electron microscopy have been used to shed light on the processes of maturation and activity of the type III secretion system and recent imaging innovations with the potential to provide better insight into the T3SS structure and function are highlighted.
Book ChapterDOI
Intracellular membrane traffic at high resolution.
Jan R.T. van Weering,Edward Brown,Thomas H. Sharp,Judith Mantell,Peter J. Cullen,Paul Verkade +5 more
TL;DR: The expansion of the technique from purely morphological analysis to cryo-immuno-EM, correlative light electron microscopy (CLEM), and 3D electron tomography is described, with a focus on the endosomal system.
Journal ArticleDOI
Intrinsically disordered regions of nucleophosmin/B23 regulate its RNA binding activity through their inter- and intra-molecular association
TL;DR: A novel mechanism by which B23 recognizes specific RNA targets is reported by which the inter- and intra-molecular interactions between the two IDRs contribute to the regulation of the RNA binding activity of CTD to control the cellular localization and functions of B23.
Journal ArticleDOI
Conventional fluorophore-free dual pH- and thermo-responsive luminescent alternating copolymer
TL;DR: In this article, a new class of traditional fluorophore-free dual pH and thermo-responsive fluorescent copolymers was synthesized through sequence-controlled copolymerization of rationally designed monomers.
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.
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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