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Fluorine NMR‐Based Screening on Cell Membrane Extracts
Author(s) -
Veronesi Marina,
Romeo Elisa,
Lambruschini Chiara,
Piomelli Daniele,
Bandiera Tiziano,
Scarpelli Rita,
Garau Gianpiero,
Dalvit Claudio
Publication year - 2014
Publication title -
chemmedchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.817
H-Index - 100
eISSN - 1860-7187
pISSN - 1860-7179
DOI - 10.1002/cmdc.201300438
Subject(s) - membrane , chemistry , enzyme , drug discovery , homogeneous , cell , cell membrane , combinatorial chemistry , biochemistry , computational biology , nuclear magnetic resonance spectroscopy , biophysics , biology , stereochemistry , physics , thermodynamics
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 number of biophysical techniques, often requiring complex set‐up, are applicable to these sample types. Here, we demonstrate the first application of n ‐fluorine atoms for biochemical screening ( n ‐FABS), a homogeneous and versatile assay based on 19 F NMR spectroscopy, to the detection of high‐ and low‐affinity inhibitors of a membrane enzyme in cell extracts and determination of their IC 50 values. Our approach can allow the discovery of novel binding fragments against targets known to be difficult to purify or where membrane‐association is required for activity. These results pave the way for future applications of the methodology to these relevant and complex biological systems.