Kinetic and structural evidence for a sequential ordered Bi Bi mechanism of catalysis by Saccharomyces cerevisiae myristoyl-CoA:protein N-myristoyltransferase

The mechanism of catalysis of Escherichia coli-derived Saccharomyces cerevisiae myristoyl-CoA: protein N-myristoyltransferase (NMT) has been characterized. Previous studies indicated that a high affinity reaction intermediate forms between NMT and myristoyl-CoA in the absence of a peptide substrate....

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Bibliographic Details
Published in:The Journal of biological chemistry 1991-05, Vol.266 (15), p.9732-9739
Main Authors: RUDNICK, D. A, MCWHERTER, C. A, ROCQUE, W. J, LENNON, P. J, GETMAN, D. P, GORDON, J. I
Format: Article
Language:English
Subjects:
Acyltransferases - metabolism
Amino Acid Sequence
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Catalysis
Enzymes and enzyme inhibitors
Fundamental and applied biological sciences. Psychology
Isoelectric Focusing
Kinetics
Molecular Sequence Data
myristoyl-CoA:protein N-myristoyltransferase
Protein Conformation
reaction mechanisms
Saccharomyces cerevisiae - enzymology
Spectrometry, Fluorescence
Spectrophotometry, Ultraviolet
Substrate Specificity
Transferases
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Summary:The mechanism of catalysis of Escherichia coli-derived Saccharomyces cerevisiae myristoyl-CoA: protein N-myristoyltransferase (NMT) has been characterized. Previous studies indicated that a high affinity reaction intermediate forms between NMT and myristoyl-CoA in the absence of a peptide substrate. This complex has been further characterized using S-(2-oxo)pentadecyl-CoA, a nonhydrolyzable myristoyl-CoA analog. Binding studies involving this analog, as well as myristoylpeptide and CoA, have indicated that the CoA moiety of the acyl substrate is retained in the acyl-NMT complex prior to peptide addition. These structural data, along with kinetic studies of myristoylpeptide and CoA product inhibition, indicate that the mechanism of catalysis of NMT is ordered Bi Bi, with myristoyl-CoA binding to NMT occurring prior to peptide binding and CoA release taking place before release of acyl peptide. Further analyses of the interactions between NMT, acyl peptide, and CoA demonstrate that NMT is able to deacylate a myristoylpeptide in the presence of CoA.
ISSN:0021-9258
1083-351X
DOI:10.1016/s0021-9258(18)92882-6