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 protein-specific glycoform information is reported. The proof-of-concept protocol developed for MUC1-specific terminal galactose/N-acetylgalactosamine (Gal/GalNAc) combines affinity binding, off-on switchable catalytic activity, and proximity catalysis to create a reactive handle for bioorthogonal labeling and imaging. Noteworthy assay features associated with LCM as compared with MOE include minimum target cell perturbation, short reaction timeframe, effectiveness as a molecular ruler, and quantitative analysis capability.A localized chemical remodeling strategy for live cell imaging of protein-specific glycoform is reported. Compared with the currently used global chemical remodeling method, namely metabolic oligosaccharide engineering, desired assay features of this approach include minimum target cell perturbation, short reaction timeframe, effectiveness as a molecular ruler, and quantitative analysis capability.
[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....
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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
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[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...
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[NMR paper] 31P NMR 2D Mapping of Creatine Kinase Forward Flux Rate in Hearts with Postinfarction Left Ventricular Remodeling in Response to Cell Therapy.
31P NMR 2D Mapping of Creatine Kinase Forward Flux Rate in Hearts with Postinfarction Left Ventricular Remodeling in Response to Cell Therapy.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--journals.plos.org-plosone-resource-img-external-pone_120x30.png Related Articles 31P NMR 2D Mapping of Creatine Kinase Forward Flux Rate in Hearts with Postinfarction Left Ventricular Remodeling in Response to Cell Therapy.
PLoS One. 2016;11(9):e0162149
Authors: Gao L, Cui W, Zhang P, Jang A, Zhu W, Zhang J
Abstract
...
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[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...
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[NMR paper] Preparation of Multiple Site-Specific Mutant Proteins for NMR Studies by PCR-Directed Cell-Free Protein Synthesis.
Preparation of Multiple Site-Specific Mutant Proteins for NMR Studies by PCR-Directed Cell-Free Protein Synthesis.
Preparation of Multiple Site-Specific Mutant Proteins for NMR Studies by PCR-Directed Cell-Free Protein Synthesis.
Methods Mol Biol. 2014;1118:169-87
Authors: Ozawa K, Qi R
Abstract
Cell-free protein synthesis (CFPS) offers a fast and inexpensive approach to selectively label proteins with isotopes that can then be detected by nuclear magnetic resonance (NMR) spectroscopy directly in the translation mixture. We describe...
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01-08-2014 11:23 AM
[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...
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[NMR paper] Remodeling of HDL by phospholipid transfer protein: demonstration of particle fusion
Remodeling of HDL by phospholipid transfer protein: demonstration of particle fusion by 1H NMR spectroscopy.
Related Articles Remodeling of HDL by phospholipid transfer protein: demonstration of particle fusion by 1H NMR spectroscopy.
Biochem Biophys Res Commun. 1998 Aug 28;249(3):910-6
Authors: Korhonen A, Jauhiainen M, Ehnholm C, Kovanen PT, Ala-Korpela M
There is evidence that phospholipid transfer protein (PLTP) can increase reverse cholesterol transport by inducing favorable subclass distribution in the high density lipoprotein (HDL)...
Localized Chemical Remodeling for Live Cell Imaging of Protein-Specific Glycoform
Linear Mode
