A fluorescence-based assay for N-myristoyltransferase activity
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Accepted version
Author(s)
Type
Journal Article
Abstract
N-myristoylation is the irreversible attachment of a C14 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.
Date Issued
2012-02-01
Date Acceptance
2011-10-05
Citation
Analytical Biochemistry, 2012, 421 (1), pp.342-344
ISSN
1096-0309
Publisher
Elsevier Masson
Start Page
342
End Page
344
Journal / Book Title
Analytical Biochemistry
Volume
421
Issue
1
Copyright Statement
© 2011, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/
Subjects
Science & Technology
Life Sciences & Biomedicine
Physical Sciences
Biochemical Research Methods
Biochemistry & Molecular Biology
Chemistry, Analytical
Chemistry
BIOCHEMICAL RESEARCH METHODS
BIOCHEMISTRY & MOLECULAR BIOLOGY
CHEMISTRY, ANALYTICAL
N-Myristoyltransferase (NMT)
Fluorescence
7-Diethylamino-3-(4-maleimido-phenyl)-4-methylcoumarin (CPM)
Coenzyme A
Screening
MYRISTOYL TRANSFERASE
IN-VIVO
INHIBITORS
COA
MEMBRANE
TARGET
GAG
Publication Status
Published