Fluorine NMR‐Based Screening on Cell Membrane Extracts. Issue 2 (11th December 2013)

• Record Type: Journal Article
• Authors: Veronesi, Marina; Romeo, Elisa; Lambruschini, Chiara; Piomelli, Daniele; Bandiera, Tiziano; Scarpelli, Rita; Garau, Gianpiero; Dalvit, Claudio
• Is Part Of: ChemMedChem. Volume 9:Issue 2(2014:Feb.)
• Page Start: 286
• Page End: 289
• Publication Date: 2013-12-11
• Subjects: Pharmaceutical chemistry -- Periodicals; 615.19005
• DOI: 10.1002/cmdc.201300438
• Languages: English
• ISSNs: 1860-7179

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <p>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 <italic>n</italic>‐fluorine atoms for biochemical screening (<italic>n</italic>‐FABS), a homogeneous and versatile assay based on ¹⁹F NMR spectroscopy, to the detection of high‐ and low‐affinity inhibitors of a membrane enzyme in cell extracts and determination of their IC₅₀ 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.</p> </abstract>