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Native Desorption Electrospray Ionization Liberates Soluble and Membrane Protein Complexes from Surfaces

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Ambrose, Stephen | Housden, Nicholas | Gupta, Kallol | Fan, Jieyuan | White, Paul | Yen, Hsin-Yung | Marcoux, Julien | Kleanthous, Colin | Hopper, Jonathan | Robinson, Carol

Edité par CCSD ; Wiley-VCH Verlag -

International audience. Mass spectrometry (MS) applications for intact protein complexes typically require electrospray(ES) ionization and have not been achieved via direct desorption from surfaces.D esorption ES ionization (DESI) MS has however transformed the study of tissue surfaces through release and characterisation of small molecules.Motivated by the desire to screen for ligand binding to intact protein complexes we report the development of an ative DESI platform. By establishing conditions that preserve non-covalent interactions we exploit the surface to capture ar apid turnover enzyme-substrate complex and to optimise detergents for membrane protein study.W ed emonstrate binding of lipids and drugs to membrane proteins deposited on surfaces and selectivity from am ix of related agonists for specific binding to aG PCR. Overall therefore we introduce this native DESI platform with the potential for high-throughput ligand screening of some of the most challenging drug targets including GPCRs.

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