Details

Autor(en) / Beteiligte

• Goncalves, Victor
• Brannigan, James A.
• Thinon, Emmanuelle
• Olaleye, Tayo O.
• Serwa, Remigiusz
• Lanzarone, Salvatore
• Wilkinson, Anthony J.
• Tate, Edward W.
• Leatherbarrow, Robin J.

Titel

A fluorescence-based assay for N-myristoyltransferase activity

Ist Teil von

• Analytical biochemistry, 2012-02, Vol.421 (1), p.342-344

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2012

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• N-myristoylation is the irreversible attachment of a C 14 fatty acid, myristic acid, to the N-terminal glycine of a protein via formation of an amide bond. This modification is catalyzed by myristoyl–coenzyme A (CoA):protein N-myristoyltransferase (NMT), an enzyme ubiquitous in eukaryotes that is up-regulated in several cancers. Here we report a sensitive fluorescence-based assay to study the enzymatic activity of human NMT1 and NMT2 based on detection of CoA by 7-diethylamino-3-(4-maleimido-phenyl)-4-methylcoumarin. We also describe expression and characterization of NMT1 and NMT2 and assay validation with small molecule inhibitors. This assay should be broadly applicable to NMTs from a range of organisms.