Document Detail


A fluorescence-based assay for N-myristoyltransferase activity.
MedLine Citation:
PMID:  22051857     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Victor Goncalves; James A Brannigan; Emmanuelle Thinon; Tayo O Olaleye; Remigiusz Serwa; Salvatore Lanzarone; Anthony J Wilkinson; Edward W Tate; Robin J Leatherbarrow
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Validation Studies     Date:  2011-10-14
Journal Detail:
Title:  Analytical biochemistry     Volume:  421     ISSN:  1096-0309     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2012 Feb 
Date Detail:
Created Date:  2012-01-16     Completed Date:  2012-05-24     Revised Date:  2014-03-19    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  342-4     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 Elsevier Inc. All rights reserved.
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MeSH Terms
Descriptor/Qualifier:
Acyltransferases / analysis*,  antagonists & inhibitors,  genetics,  metabolism
Coenzyme A
Coumarins
Fluorescence
Fluorescent Dyes
Humans
Kinetics
Myristic Acids / metabolism
Protein Processing, Post-Translational
Grant Support
ID/Acronym/Agency:
087792//Wellcome Trust; 087792//Wellcome Trust; 10711//Cancer Research UK; BB/D02014X/1//Biotechnology and Biological Sciences Research Council; C29637/A10711//Cancer Research UK; G0900278//Medical Research Council; G0900278//Medical Research Council
Chemical
Reg. No./Substance:
0/Coumarins; 0/Fluorescent Dyes; 0/Myristic Acids; 76877-33-3/N-(4-(7-diethylamino-4-methylcoumarin-3-yl)phenyl)maleimide; EC 2.3.-/Acyltransferases; EC 2.3.1.97/glycylpeptide N-tetradecanoyltransferase; SAA04E81UX/Coenzyme A
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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