Comparison of the acyl chain specificities of human myristoyl-CoA synthetase and human myristoyl-CoA:protein N-myristoyltransferase.

Kishore, Nandini S.; Wood, David; Mehta, Pramod P.; Wade, Arlene C.; Lu, Taotao; Gokel, George W.; Gordon, Jeffrey I.

doi:10.1016/s0021-9258(18)53479-7

Cited by 69 publications

(28 citation statements)

References 74 publications

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“…The preference for glycine is well understood from structural studies as the side chain of other amino acids are not tolerated in the active site because of steric reasons . However, recent studies demonstrated that human NMT1/2 can also catalyze the myristoylation of lysine 3 side chain of ARF6 protein. , Previously, the activity of partially purified human NMT was tested on a variety of acyl-CoA molecules ranging from C7 to C16, and myristoyl-CoA is shown to be the most preferred substrate . However, shorter acyl-CoA molecules were not tested.…”

Section: Resultsmentioning

confidence: 99%

“…5,6 Previously, the activity of partially purified human NMT was tested on a variety of acyl-CoA molecules ranging from C7 to C16, and myristoyl-CoA is shown to be the most preferred substrate. 7 However, shorter acyl-CoA molecules were not tested. Surprisingly, when we tested recombinant h u m a n N M T 1 w i t h A R F 6 N -t e r m i n a l p e p t i d e (GKVLSKIFWW), we found that acetyl-CoA can also be used as a substrate, leading to the formation of an acetyl peptide product (Figure 1).…”

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confidence: 99%

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Binding Affinity Determines Substrate Specificity and Enables Discovery of Substrates for N-Myristoyltransferases

Kosciuk

Yang

et al. 2021

ACS Catal.

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Kinetic parameters ( k cat and K m ) derived from the Michaelis–Menten equation are widely used to characterize enzymes. k cat / K m is considered the catalytic efficiency or substrate specificity of an enzyme toward its substrate. N-Myristoyltransferases (NMTs) catalyze the N-terminal glycine myristoylation of numerous eukaryotic proteins. Surprisingly, we find that in vitro human NMT1 can accept acetyl-CoA and catalyze acetylation with k cat and K m values similar to that of myristoylation. However, when both acetyl-CoA and myristoyl-CoA are present in the reaction, NMT1 catalyzes almost exclusively myristoylation. This phenomenon is caused by the dramatically different binding affinities of NMT1 for myristoyl-CoA and acetyl-CoA (estimated K d of 14.7 nM and 10.1 μM, respectively). When both are present, NMT1 is essentially entirely bound by myristoyl-CoA and thus catalyzes myristoylation exclusively. The NMT1 example highlights the crucial role of binding affinity in determining the substrate specificity of enzymes, which in contrast to the traditionally held view in enzymology that the substrate specificity is defined by k cat / K m values. This understanding readily explains the vast biological literature showing the coimmunoprecipitation of enzyme–substrate pairs for enzymes that catalyzes protein post-translational modifications (PTM), including phosphorylation, acetylation, and ubiquitination. Furthermore, this understanding allows the discovery of substrate proteins by identifying the interacting proteins of PTM enzymes, which we demonstrate by identifying three previously unknown substrate proteins (LRATD1, LRATD2, and ERICH5) of human NMT1/2 by mining available interactome data.

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

Binding Affinity Determines Substrate Specificity and Enables Discovery of Substrates for N-Myristoyltransferases

Kosciuk

Yang

et al. 2021

ACS Catal.

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“…Several studies have shown that NMT is a potential target for antifungal and antiparasite ,, drugs because it is indispensable for the growth and viability of fungal and parasitic organisms. Moreover, compared with the myristoyl-CoA binding site, the peptide binding pocket of NMT is less well-conserved across species . The pocket can therefore be targeted for the development of selective NMT inhibitors.…”

Section: N-terminal Glycine Myristoylationmentioning

confidence: 99%

“…Moreover, compared with the myristoyl-CoA binding site, the peptide binding pocket of NMT is less well-conserved across species. 339 The pocket can therefore be targeted for the development of selective NMT inhibitors. Several series of inhibitors (Figure 17) from high-throughput screening have been reported for NMTs in humans, 340 parasites (P. falciparum, Leishmania sp., T. brucei), 338,341−344 and fungi.…”

Section: Chemical Reviewsmentioning

confidence: 99%

Protein Lipidation: Occurrence, Mechanisms, Biological Functions, and Enabling Technologies

et al. 2018

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Protein lipidation, including cysteine prenylation, N-terminal glycine myristoylation, cysteine palmitoylation, and serine and lysine fatty acylation, occurs in many proteins in eukaryotic cells and regulates numerous biological pathways, such as membrane trafficking, protein secretion, signal transduction, and apoptosis. We provide a comprehensive review of protein lipidation, including descriptions of proteins known to be modified and the functions of the modifications, the enzymes that control them, and the tools and technologies developed to study them. We also highlight key questions about protein lipidation that remain to be answered, the challenges associated with answering such questions, and possible solutions to overcome these challenges.

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“…MyrCoA analogs can be obtained in vitro from free CoA and any FA derivative using either chemical or enzymatic approaches [151][152][153]. From studies of such compounds, it appears that NMT can also efficiently accommodate C10 to C15 chains [154,155] and appears to tolerate variations at the extreme side of the acyl chain beyond C5/6 [156][157][158]. Nevertheless, C14:0 is considered the major acyl form used by NMT.…”

Section: Main Structural and Biochemical Features Of Nmtsmentioning

confidence: 99%

Mapping the myristoylome through a complete understanding of protein myristoylation biochemistry

Giglione

Meinnel

2022

Progress in Lipid Research

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Comparison of the acyl chain specificities of human myristoyl-CoA synthetase and human myristoyl-CoA:protein N-myristoyltransferase.

Cited by 69 publications

References 74 publications

Binding Affinity Determines Substrate Specificity and Enables Discovery of Substrates for N-Myristoyltransferases

Binding Affinity Determines Substrate Specificity and Enables Discovery of Substrates for N-Myristoyltransferases

Protein Lipidation: Occurrence, Mechanisms, Biological Functions, and Enabling Technologies

Mapping the myristoylome through a complete understanding of protein myristoylation biochemistry

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