The substrate- or cofactor-based fluorine NMR screening, also known as n-FABS (n fluorine atoms for biochemical screening), represents a powerful method for performing a direct functional assay in the search of inhibitors or enhancers of an enzymatic reaction. Although it suffers from the intrinsic low sensitivity compared to other biophysical techniques usually applied in functional assays, it has some distinctive features that makes it appealing for tackling complex chemical and biological systems. Its strengths are represented by the easy set-up, robustness, flexibility, lack of signal interference and rich information content resulting in the identification of bona fide inhibitors and reliable determination of their inhibitory strength. The versatility of the n-FABS allows its application to either purified enzymes, cell lysates or intact living cells. The principles, along with theoretical, technical and practical aspects, of the methodology are discussed. Furthermore, several applications of the technique to pharmaceutical projects are presented.
[NMR paper] Recent progress of in-cell NMR of nucleic acids in living human cells.
Recent progress of in-cell NMR of nucleic acids in living human cells.
Related Articles Recent progress of in-cell NMR of nucleic acids in living human cells.
Biophys Rev. 2020 Mar 06;:
Authors: Yamaoki Y, Nagata T, Sakamoto T, Katahira M
Abstract
The inside of living cells is highly crowded with biological macromolecules. It has long been considered that the properties of nucleic acids and proteins, such as their structures, dynamics, interactions, and enzymatic activities, in intracellular environments are different from those...
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03-09-2020 01:12 PM
[NMR paper] The first successful observation of in-cell NMR signals of DNA and RNA in living human cells.
The first successful observation of in-cell NMR signals of DNA and RNA in living human cells.
Related Articles The first successful observation of in-cell NMR signals of DNA and RNA in living human cells.
Phys Chem Chem Phys. 2017 Oct 12;:
Authors: Yamaoki Y, Kiyoishi A, Miyake M, Kano F, Murata M, Nagata T, Katahira M
Abstract
In order to understand intracellular biological events, information on the structure, dynamics and interaction of proteins and nucleic acids in living cells is of crucial importance. In-cell NMR is a...
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10-14-2017 02:04 AM
[NMR paper] Weak Intermolecular Hydrogen Bonds with Fluorine: Detection and Implications for Enzymatic/Chemical Reactions, Chemical Properties, and Ligand/Protein Fluorine NMR Screening.
Weak Intermolecular Hydrogen Bonds with Fluorine: Detection and Implications for Enzymatic/Chemical Reactions, Chemical Properties, and Ligand/Protein Fluorine NMR Screening.
Related Articles Weak Intermolecular Hydrogen Bonds with Fluorine: Detection and Implications for Enzymatic/Chemical Reactions, Chemical Properties, and Ligand/Protein Fluorine NMR Screening.
Chemistry. 2016 Apr 26;
Authors: Dalvit C, Vulpetti A
Abstract
It is known that strong hydrogen-bonding interactions play an important role in many chemical and...
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04-27-2016 01:51 PM
[NMR paper] Fluorine NMR-based assay in living mammalian cells.
Fluorine NMR-based assay in living mammalian cells.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Fluorine NMR-based assay in living mammalian cells.
Anal Biochem. 2015 Dec 11;
Authors: Veronesi M, Giacomina F, Romeo E, Castellani B, Ottonello G, Lambruschini C, Garau G, Scarpelli R, Bandiera T, Piomelli D, Dalvit C
Abstract
Nuclear Magnetic Resonance (NMR)-based screening has been recognized as a powerful approach for the identification...
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12-28-2015 12:26 AM
Affinity screening using competitive binding with fluorine-19 hyperpolarized ligands
From The DNP-NMR Blog:
Affinity screening using competitive binding with fluorine-19 hyperpolarized ligands
Kim, Y. and C. Hilty, Affinity screening using competitive binding with fluorine-19 hyperpolarized ligands. Angew Chem Int Ed Engl, 2015. 54(16): p. 4941-4.
http://www.ncbi.nlm.nih.gov/pubmed/25703090
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10-08-2015 12:56 AM
[NMR paper] Functional Binding Surface of a ?-Hairpin VEGF Receptor Targeting Peptide Determined by NMR Spectroscopy in Living Cells.
Functional Binding Surface of a ?-Hairpin VEGF Receptor Targeting Peptide Determined by NMR Spectroscopy in Living Cells.
Functional Binding Surface of a ?-Hairpin VEGF Receptor Targeting Peptide Determined by NMR Spectroscopy in Living Cells.
Chemistry. 2014 Nov 6;
Authors: Diana D, Russomanno A, Rosa LD, Di Stasi R, Capasso D, Di Gaetano S, Romanelli A, Russo L, D'Andrea LD, Fattorusso R
Abstract
In this study, the functional interaction of HPLW peptide with VEGFR2 (Vascular Endothelial Growth Factor Receptor 2) was determined...
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11-08-2014 12:43 PM
[NMR paper] Fluorine NMR-Based Screening on Cell Membrane Extracts.
Fluorine NMR-Based Screening on Cell Membrane Extracts.
Related Articles Fluorine NMR-Based Screening on Cell Membrane Extracts.
ChemMedChem. 2013 Dec 11;
Authors: Veronesi M, Romeo E, Lambruschini C, Piomelli D, Bandiera T, Scarpelli R, Garau G, Dalvit C
Abstract
The possibility of measuring the action of inhibitors of specific enzymatic reactions in intact cells, cell lysates or membrane preparations represents a major advance in the lead discovery process. Despite the relevance of assaying in physiological conditions, only a small...
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12-18-2013 04:00 PM
[NMR paper] 1H-NMR investigation of the oxygenation of hemoglobin in intact human red blood cells
1H-NMR investigation of the oxygenation of hemoglobin in intact human red blood cells.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-cellhub.gif http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles 1H-NMR investigation of the oxygenation of hemoglobin in intact human red blood cells.
Biophys J. 1995 Feb;68(2):681-93
Authors: Fetler BK, Simplaceanu V, Ho C
Using improved selective excitation methods for protein nuclear...
Fluorine NMR functional screening: from purified enzymes to human intact living cells
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