A simplified assay for the enzyme responsible for the attachment of myristic acid to the N-terminal glycine residue of proteins, myristoyl-CoA: glycylpeptide N-myristoyltransferase.
A greatly simplified assay for myristoyl-CoA:glycylpeptide N-myristoyltransferase (NMT) activity is described. The assay is based on the differential solubility of the acyl-peptides produced as a consequence of the NMT activity and yields results comparable with those obtained with the original assay described by Towler & Glaser [(1986) Proc. Natl. Acad. Sci. U.S.A. 83, 2812-2816], which requires h.p.l.c. to determine the production of the acyl-peptides. The use of the revised assay in the preliminary steps of the purification of rat brain NMT is described, and its use in determining the fatty acid-specificity of the enzyme is illustrated. The results are shown to be comparable with those obtained with the h.p.l.c.-based assay.
McIlhinney
R A
RA
Department of Pharmacology, University of Oxford, U.K.
McGlone
K
K
D000214:Acyl Coenzyme A / Q000378:metabolism; D000217:Acyltransferases / Q000032:analysis* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D003065:Coenzyme A / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005998:Glycine / Q000378:metabolism*; D008297:Male; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility
D000214:Acyl Coenzyme A; D009227:Myristic Acids; D010455:Peptides; D019814:Myristic Acid; C040349:S-tetradecanoyl-coenzyme A; D000255:Adenosine Triphosphate; D000217:Acyltransferases; C050480:glycylpeptide N-tetradecanoyltransferase; D003065:Coenzyme A; D005998:Glycine