Acylation-coupled lipophilic induction of polarisation (Acyl-cLIP): a universal assay for lipid transferase and hydrolase enzymes.

Autor:	Lanyon-Hogg T; Department of Chemistry , Imperial College London , London , W12 0BZ , UK . Email: t.lanyon-hogg@imperial.ac.uk ; Email: e.tate@imperial.ac.uk ; Tel: +44 0207 5943752 ; Tel: +44 0207 5945821., Ritzefeld M; Department of Chemistry , Imperial College London , London , W12 0BZ , UK . Email: t.lanyon-hogg@imperial.ac.uk ; Email: e.tate@imperial.ac.uk ; Tel: +44 0207 5943752 ; Tel: +44 0207 5945821., Sefer L; Division of Structural Biology , Wellcome Centre for Human Genetics , University of Oxford , Oxford , OX3 7BN , UK., Bickel JK; Department of Chemistry , Imperial College London , London , W12 0BZ , UK . Email: t.lanyon-hogg@imperial.ac.uk ; Email: e.tate@imperial.ac.uk ; Tel: +44 0207 5943752 ; Tel: +44 0207 5945821., Rudolf AF; Division of Structural Biology , Wellcome Centre for Human Genetics , University of Oxford , Oxford , OX3 7BN , UK., Panyain N; Department of Chemistry , Imperial College London , London , W12 0BZ , UK . Email: t.lanyon-hogg@imperial.ac.uk ; Email: e.tate@imperial.ac.uk ; Tel: +44 0207 5943752 ; Tel: +44 0207 5945821., Bineva-Todd G; The Francis Crick Institute , London NW1 1AT , UK., Ocasio CA; The Francis Crick Institute , London NW1 1AT , UK., O'Reilly N; The Francis Crick Institute , London NW1 1AT , UK., Siebold C; Division of Structural Biology , Wellcome Centre for Human Genetics , University of Oxford , Oxford , OX3 7BN , UK., Magee AI; Molecular Medicine Section , National Heart & Lung Institute , Imperial College London , London , SW7 2AZ , UK., Tate EW; Department of Chemistry , Imperial College London , London , W12 0BZ , UK . Email: t.lanyon-hogg@imperial.ac.uk ; Email: e.tate@imperial.ac.uk ; Tel: +44 0207 5943752 ; Tel: +44 0207 5945821.
Jazyk:	angličtina
Zdroj:	Chemical science [Chem Sci] 2019 Jul 01; Vol. 10 (39), pp. 8995-9000. Date of Electronic Publication: 2019 Jul 01 (Print Publication: 2019).
DOI:	10.1039/c9sc01785b
Abstrakt:	Posttranslational attachment of lipids to proteins is important for many cellular functions, and the enzymes responsible for these modifications are implicated in many diseases, from cancer to neurodegeneration. Lipid transferases and hydrolases are increasingly tractable therapeutic targets, but present unique challenges for high-throughput biochemical enzyme assays which hinder development of new inhibitors. We present Acylation-coupled Lipophilic Induction of Polarisation (Acyl-cLIP) as the first universally applicable biochemical lipidation assay, exploiting the hydrophobic nature of lipidated peptides to drive a polarised fluorescence readout. Acyl-cLIP allows sensitive, accurate, real-time measurement of S - or N -palmitoylation, N -myristoylation, S -farnesylation or S -geranylgeranylation. Furthermore, it is applicable to transfer and hydrolysis reactions, and we demonstrate its extension to a high-throughput screening format. We anticipate that Acyl-cLIP will greatly expedite future drug discovery efforts against these challenging targets. (This journal is © The Royal Society of Chemistry 2019.)
Databáze:	MEDLINE
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