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
Dynamic fluorescence depolarization: a powerful tool to explore protein folding on the ribosome.
TL;DR: The theory of fluorescence depolarization and its exciting applications to the study of the dynamics of nascent proteins in the cellular environment are discussed.
Journal ArticleDOI
CrAsH-quantum dot nanohybrids for smart targeting of proteins.
Emilie Genin,Olivier Carion,Benoit Mahler,Benoit Dubertret,Nathalie J. Arhel,Pierre Charneau,Eric Doris,Charles Mioskowski +7 more
TL;DR: Smart nanohybrids prepared by conjugation of CrAsH to hydrosoluble and biocompatible quantum dots (QDs) were shown to bind efficiently and selectively to Cys-tagged proteins to allow extended monitoring of the target protein.
Patent
Modified leptin polypeptides and their uses
Vadim Kraynov,Anna-Maria A. Hays Putnam,Nick Knudsen,Jason Pinkstaff,Heather Myler,Lorraine Sullivan +5 more
TL;DR: In this paper, the problem of providing leptin polypeptides modified with at least one not naturally encoded amino acid is solved by linking the amino acid to a water-soluble polymer.
Journal ArticleDOI
Remodelling epithelial tubes through cell rearrangements: from cells to molecules.
Marc Neumann,Markus Affolter +1 more
TL;DR: Recent progress in understanding cell rearrangements during epithelial tube remodelling is described and several models that might account for the developmental control of the spatial dynamics of adherens junctions are discussed.
Journal ArticleDOI
Intracellular protein labeling with prodrug-like probes using a mutant β-lactamase tag
TL;DR: This prodrug-based probe design strategy for BL-tags provides a simple experimental procedure with application to cellular studies with the additional advantage of reduced stress to living cells.
References
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Journal ArticleDOI
The green fluorescent protein
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Journal ArticleDOI
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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’.
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
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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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