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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Patent
Compositions and methods for determining interactions of mitochondrial components, and for identifying agents that alter such interactions
Anne N. Murphy,William Clevenger,Sandra E. Wiley,Alexander Y. Andreyev,Luciano G. Frigeri,Gonul Velicelebi,Robert E. Davis +6 more
TL;DR: In this article, compositions and methods for identifying agents that alter the mitochondrial membrane permeability transition were provided, and the screening methods generally detect agents that altered the interaction between the mitochondrial adenine nucleotide translocator and cyclophilin D.
Journal ArticleDOI
In Situ Construction of Protein-Based Semisynthetic Biosensors
TL;DR: Recent efforts regarding the in situ construction of protein-based semisynthetic biosensors are discussed and their uses in the visualization and quantification of biomolecules and events in multimolecular and crowded cellular systems are highlighted.
Journal ArticleDOI
Bacterial Vivisection: How Fluorescence-Based Imaging Techniques Shed a Light on the Inner Workings of Bacteria.
Alexander Cambré,Abram Aertsen +1 more
TL;DR: The current state of the art in fluorescence-based vivisection of bacteria is outlined and an overview of important case studies to exemplify how to use or combine different strategies to gain detailed information on the cell’s physiology is provided.
Journal ArticleDOI
Target-protein-selective inactivation and labelling using an oxidative catalyst
TL;DR: This review focuses on two approaches of selective inactivation of protein functions and selective protein labelling using oxidative SET catalysts that can catalyze oxidative SET reactions in a protein mixture under intracellular conditions.
Journal ArticleDOI
Fluorosomes: Fluorescent Virus-Like Nanoparticles that Represent a Convenient Tool to Visualize Receptor-Ligand Interactions
TL;DR: An overview on virus particle assembly, chemical and recombinant methods of virus particle labeling and examples on how FS can be applied as sensors to monitor receptor-ligand interactions on leukocytes are given.
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
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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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