Acylation-coupled lipophilic induction of polarisation (Acyl-cLIP): a universal assay for lipid transferase and hydrolase enzymes. Issue 39 (14th August 2019)

• Record Type: Journal Article
• Authors: Lanyon-Hogg, Thomas; Ritzefeld, Markus; Sefer, Lea; Bickel, Jasmine K.; Rudolf, Amalie F.; Panyain, Nattawadee; Bineva-Todd, Ganka; Ocasio, Cory A.; O'Reilly, Nicola; Siebold, Christian; Magee, Anthony I.; Tate, Edward W.
• Is Part Of: Chemical science. Volume 10:Issue 39(2019)
• Page Start: 8995
• Page End: 9000
• Publication Date: 2019-08-14
• Subjects: Chemistry -- Periodicals; 540.5
• DOI: 10.1039/c9sc01785b
• Languages: English
• ISSNs: 2041-6520

Abstract

Abstract : A highly accurate and versatile fluorescence polarisation assay for any enzyme adding or removing lipid posttranslational modifications, with the potential to accelerate drug discovery against these targets. Abstract : 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.