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
Rational design of boron dipyrromethene (BODIPY)-based photobleaching- resistant fluorophores applicable to a protein dynamics study
Toru Komatsu,Daihi Oushiki,Aoi Takeda,Masaki Miyamura,Tasuku Ueno,Takuya Terai,Kenjiro Hanaoka,Yasuteru Urano,Tomoko Mineno,Tetsuo Nagano +9 more
TL;DR: It is found that the photobleaching rate is influenced by the electron-withdrawing capacity of the substituents of boron dipyrromethene derivatives, and the utility of one of these fluorophores, 2,6-diCO(2)R-BDP, for visualizing EGF receptor dynamics in cells expressing an SNAP-tagged EGF receptors is confirmed.
Book ChapterDOI
Protein labeling with FlAsH and ReAsH.
TL;DR: The combination of a small genetically encoded peptide tag with a small molecule detection reagent makes this technology particularly suitable for the investigation of biochemical changes in living cells that are difficult to approach with fluorescent proteins as molecular tags.
Journal ArticleDOI
Methylated and thiolated arsenic species for environmental and health research - A review on synthesis and characterization
William R. Cullen,Qingqing Liu,Xiufen Lu,Anthony McKnight-Whitford,Hanyong Peng,Aleksandra Popowich,Xiaowen Yan,Qi Zhang,Michael Fricke,Hongsui Sun,X. Chris Le +10 more
TL;DR: On the basis of reaction yield, ease of synthesis and purification of product, safety considerations, and the experience, this work recommends a method for the synthesis of each of these arsenic compounds.
Journal ArticleDOI
Protein labeling with fluorogenic probes for no-wash live-cell imaging of proteins.
Yuichiro Hori,Kazuya Kikuchi +1 more
TL;DR: The design strategy of the probes and the advances in fluorogenic protein labeling systems are described, allowing rapid detection of proteins in living cells with high signal-to-noise ratio.
Journal ArticleDOI
Specifically and wash-free labeling of SNAP-tag fused proteins with a hybrid sensor to monitor local micro-viscosity
TL;DR: A hybrid sensor BDP-V BG is developed by connecting a viscosity-sensitive boron-dipyrromethene (BODIPY) molecular rotor to O6-benzylguanine (BG) for specific detection of local micro-viscosity of SNAP-tag fused proteins and achieves high specific labeling of cells expressing two SNAP- tag fused proteins.
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
TL;DR: The reliability and sensitivity of the test have been increased to the point where it can in many cases replace the [3H]thymidine uptake assay to measure cell proliferation or survival in growth factor or cytotoxicity assays.
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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