The development of bioorthogonal approaches for labeling of endogenous proteins under the multimolecular crowding conditions of live cells is highly desirable for the analysis and engineering of proteins without using genetic manipulation. Herein, we report N-sulfonyl pyridone (SP) as a new reactive group for protein sulfonylation. The ligand-directed SP chemistry was able to modify not only purified proteins in vitro, but also endogenous ones on the surface of and inside live cells selectively and rapidly, which allowed to convert endogenous proteins to FRET-based biosensors in situ.
[NMR paper] Proximity-triggered covalent stabilization of low-affinity protein complexes in vitro and in vivo
Proximity-triggered covalent stabilization of low-affinity protein complexes in vitro and in vivo
The characterization of low-affinity protein complexes is challenging due to their dynamic nature. Here we present a method to stabilize transient protein complexes in vivo by generating a covalent and conformationally flexible bridge between the interaction partners. A highly active pyrrolysyl tRNA synthetase mutant directs the incorporation of unnatural amino acids bearing bromoalkyl moieties (BrCnK) into proteins. We demonstrate for the first time that low-affinity protein complexes...
nmrlearner
Journal club
0
09-28-2017 07:57 PM
[NMR paper] Localized Chemical Remodeling for Live Cell Imaging of Protein-Specific Glycoform
Localized Chemical Remodeling for Live Cell Imaging of Protein-Specific Glycoform
Live cell imaging of protein-specific glycoforms is important for the elucidation of glycosylation mechanisms and identification of disease states. The currently used metabolic oligosaccharide engineering (MOE) technology permits routinely global chemical remodeling (GCM) for carbohydrate site of interest, but can exert unnecessary whole-cell scale perturbation and generate unpredictable metabolic efficiency issue. A localized chemical remodeling (LCM) strategy for efficient and reliable access to...
nmrlearner
Journal club
0
06-13-2017 06:55 AM
[NMR paper] A general mechanism of photoconversion of green-to-red fluorescent proteins based on blue and infrared light reduces phototoxicity in live-cell single-molecule imaging
A general mechanism of photoconversion of green-to-red fluorescent proteins based on blue and infrared light reduces phototoxicity in live-cell single-molecule imaging
Photoconversion of fluorescent proteins by blue and complementary near-infrared light, termed primed conversion (PC), is a mechanism recently discovered for Dendra2. We demonstrate that controlling the conformation of arginine at residue 66 by threonine at residue 69 of fluorescent proteins from Anthozoan families (Dendra2, mMaple, Eos, mKikGR, pcDronpa protein families) represents a general route to facilitate PC....
nmrlearner
Journal club
0
06-02-2017 08:33 PM
The Cation-? Interaction Enables a Halo-TagFluorogenic Probe for Fast No-Wash Live Cell Imaging and Gel-FreeProtein Quantification
The Cation-? Interaction Enables a Halo-TagFluorogenic Probe for Fast No-Wash Live Cell Imaging and Gel-FreeProtein Quantification
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.7b00056/20170308/images/medium/bi-2017-00056j_0007.gif
Biochemistry
DOI: 10.1021/acs.biochem.7b00056
http://feeds.feedburner.com/~ff/acs/bichaw?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/bichaw/~4/K_EHWBvChxU
More...
nmrlearner
Journal club
0
03-14-2017 08:16 AM
[NMR paper] Insights into the Binding of Cyclic RGD Peptidomimetics to ?5?1 Integrin by using Live-Cell NMR And Computational Studies.
Insights into the Binding of Cyclic RGD Peptidomimetics to ?5?1 Integrin by using Live-Cell NMR And Computational Studies.
Related Articles Insights into the Binding of Cyclic RGD Peptidomimetics to ?5?1 Integrin by using Live-Cell NMR And Computational Studies.
ChemistryOpen. 2017 Feb;6(1):128-136
Authors: Guzzetti I, Civera M, Vasile F, Arosio D, Tringali C, Piarulli U, Gennari C, Pignataro L, Belvisi L, Potenza D
Abstract
The interaction of a small library of cyclic DKP-RGD peptidomimetics with ?5?1 integrin has been...
nmrlearner
Journal club
0
02-09-2017 12:19 PM
[NMR paper] Live Cell NMR.
Live Cell NMR.
Related Articles Live Cell NMR.
Annu Rev Biophys. 2014 May 6;43:171-192
Authors: Freedberg DI, Selenko P
Abstract
Ever since scientists realized that cells are the basic building blocks of all life, they have been developing tools to look inside them to reveal the architectures and mechanisms that define their biological functions. Whereas "looking into cells" is typically said in reference to optical microscopy, high-resolution in-cell and on-cell nuclear magnetic resonance (NMR) spectroscopy is a powerful method...
[NMR paper] Quantitative comparison of protein dynamics in live cells and in vitro by in-cell (19)F-NMR.
Quantitative comparison of protein dynamics in live cells and in vitro by in-cell (19)F-NMR.
Quantitative comparison of protein dynamics in live cells and in vitro by in-cell (19)F-NMR.
Chem Commun (Camb). 2013 Feb 26;
Authors: Takaoka Y, Kioi Y, Morito A, Otani J, Arita K, Ashihara E, Ariyoshi M, Tochio H, Shirakawa M, Hamachi I
Abstract
Here we describe how a (19)F-probe incorporated into an endogenous protein by a chemical biology method revealed protein dynamics. By explicit determination of ligand-bound and unbound structures with...
Live-cell Protein Sulfonylation Based on Proximity-driven N-Sulfonyl Pyridone Chemistry
Linear Mode
