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
Chemistry for Covalent Modification of Endogenous/Native Proteins: From Test Tubes to Complex Biological Systems
Tomonori Tamura,Itaru Hamachi +1 more
TL;DR: A variety of promising strategies have appeared recently that address this grand challenge in chemical biology and yield native protein-based well-defined bioconjugations, specific labeling of endogenous proteins in various biological crude milieus, and the establishment of chemical proteomics as a new research area in protein science.
Journal ArticleDOI
Calcium Green FlAsH as a genetically targeted small-molecule calcium indicator
Oded Tour,Stephen R. Adams,Rex Kerr,Rex Kerr,Rex Kerr,Rene M Meijer,Terrence J. Sejnowski,Richard W. Tsien,Roger Y. Tsien +8 more
TL;DR: Intracellular Ca(2+) regulates numerous proteins and cellular functions and can vary substantially over submicron and submillisecond scales, so precisely localized fast detection is desirable, and CaGF can report highly localized, rapid [Ca(2+)] dynamics.
Journal ArticleDOI
Genetic Incorporation of a Small, Environmentally Sensitive, Fluorescent Probe into Proteins in Saccharomyces cerevisiae
TL;DR: It is reported that the fluorescent amino acid, 3-acetylnaphthalen-2-ylamino)-2-aminopropanoic acid (Anap), can be genetically incorporated into proteins in yeast with excellent selectivity and efficiency by means of an orthogonal tRNA/aminoacyl-tRNA synthetase pair.
Journal ArticleDOI
Non-genetic engineering of cells for drug delivery and cell-based therapy.
TL;DR: Non-genetic cell engineering of cells has been applied in delivering therapeutics to tissues, homing of cells to the bone marrow or inflammatory tissues, cancer imaging, immunotherapy, and remotely controlling cellular functions.
Journal ArticleDOI
Characterization of three classes of membrane proteins involved in fungal azole resistance by functional hyperexpression in Saccharomyces cerevisiae.
Erwin Lamping,Brian C. Monk,Kyoko Niimi,Ann R. Holmes,Sarah Tsao,Koichi Tanabe,Masakazu Niimi,Yoshimasa Uehara,Richard D. Cannon +8 more
TL;DR: The use of a modified membrane protein hyperexpression system to characterize three classes of fungal membrane proteins that contribute to the drug resistance phenotypes of five pathogenic fungi and to express human P glycoprotein.
References
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Journal ArticleDOI
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Journal ArticleDOI
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Journal ArticleDOI
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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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
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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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