Abstract
N-myristoyltransferase (NMT) is an essential eukaryotic enzyme that catalyzes the transfer of myristate to the NH2-terminal glycine residue of a number of important proteins of diverse function. Little is known about the control and regulation of NMT in higher eukaryotes. Bovine spleen N-myristoyltransferase has been purified and characterized [Raju, RVS, Kalra J & Sharma RK (1994) J Biol Chem 269:12080–12083]. The activation of bovine spleen NMT with thiol reducing compounds, and its inhibition by the oxidizing agent sodium iodate, suggest a role for oxidation/reduction in NMT regulation. Available knowledge concerning coenzyme A (CoA), the thiol in the cell, indicated that the agents tested on NMT could also reduce or oxidize CoA. The studies suggested that reduced CoA is the key regulator of NMT activity, while oxidized CoA did not allow NMT to promote myristoylation. Further, the process of myristoylation and demyristoylation may be governed by NMT, depending on the differential concentration of CoA. The process of demyristoylation could be blocked by excess CoA. We therefore hypothesize that the initial event in the regulation of NMT is an increase in cellular CoA concentration which could be coupled to an increase in protein myristoylation. Once the CoA concentration in the cell decreases due to oxidation, the demyristoylation process would be operative.
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Abbreviations
- NMT:
-
N-myristoyl CoA:protein N-myristoyltransferase
- hNMT:
-
human NMT
- YNMT:
-
yeast NMT
- DTNB:
-
N-5′5 dithiobis(2-nitrobenzoic acid)
- DTT:
-
dithiothretol
- 2-ME:
-
2-mercaptoethanol
References
Johnson DR, Bhatnagar RS, Knoll LJ, Gordon JI: Genetic and biochemical studies of protein N-myristoylation. Ann Rev Biochem 63: 869–914, 1994
Raju RVS, Magnuson BA, Sharma RK: Mammalian N-myristoyl CoA:protein N-myristoyltransferase. Mol Cell Biochem 149/150: 191–202, 1995
McLaughlin S, Aderem A: The myristoyl-electrostatic switch: a modulator of reversible protein-membrane interaction. TIBS 235: 272–276, 1995
Duronio RJ, Reed SI, Gordon JI: Mutations of human myristoyl-CoA:protein N-myristoyltransferase cause temperature-sensitive myristoyl group auxotrophy in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 89: 4129–4133, 1992
Johnson DR, Duronio RJ, Langner CA, Rudnick DA, Gordon JI: Genetic and biochemical studies of a mutant Saccharomyces cerevisiae myristoyl-CoA-protein N-myristoyltransferase nmt72Leu99 → Pro, that produce temperature -sensitive myristoyl group auxotrophy. J Biol Chem 268:483–494, 1993
Bryant ML, Heuckeroth RO, Kimata TJ, Ratner L, Gordon JI: Replication of human immunodeficiency virus-1 and moloney murine leukaemia virus is inhibited by different heteroatomcontaining analogs of a myristic acid. Proc Natl Acad Sci USA 86: 8655–8659, 1989
Gallay P, Swingler S, Aiken C, Trono D: HIV-1 Infection of nondividing cells: C-terminal tyrosine phosphorylation of the viral matrix protein is a key regulator. Cell 80: 379–388, 1995
Tashiro A, Shoji S, Kubota Y: Antimyristoylation of the gag proteins in the human immunodeficiency virus infected cells with N-myristoyl glycinal diethyl acetal resulted in inhibition of virus production. Biochem Biophys Res Commun 165: 1145–1154, 1989
Saermark T, Kleinschmidt A, Wulff AM, Andreassen H, Magee AI: Characterization of N-myristoyl transferase inhibitors and their effect on HIV release. AIDS 5: 951–958, 1991
Magnuson BA, Raju RVS, Moyana TN, Sharma RK: Increased N-myristoyltransferase activity observed in rat and human colonic tumors. J Natl Cancer Inst 87: 1630–1635, 1995
King MJ, Sharma RK: Differential activation of bovine brain myristoyltransferase(s) by a cytosolic activator. Biochem Biophys Res Commun 212: 580–588, 1995
King MJ, Sharma RK: Identification, purification and characterization of a membraneassociated N-myristoyltransferase inhibitor protein from bovine brain. Biochem J 291: 635–639, 1993
Merrifield RB: Solid phase synthesis, Science 232: 341–347, 1986
King MJ, Sharma RK: N-myristoyltransferase assay using phosphocellulose paper binding, Anal Biochem 119: 149–153, 1991
Raju RVS, Kalra J, Sharma RK: Purification and properties of bovine spleen N-myristoyl CoA:proteinN-myristoyltransferase. J Biol Chem 269:12080–12083, 1994
Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle protein-dye binding. Anal Biochem 72: 248–254, 1976
Ellman GL: Tissue sulthydryl groups. Arch Biochem Biophys 82: 70–77, 1959
CecilR, McPhee JR: The sulfur chemistry of proteins. Adv Protein Chem 14:255–389, 1959
Raju RVS, Sharma RK: Myristoyl CoA:protein N-myristoyltransferase: Subcellular localization, activation and kinetic behaviour in the presence of organic solvents, Biochem Biophys Res Commun 208: 617–623, 1995
Towler DA, Adams SP, Eubanks SR, Towery DS, Jackson-Machelski E, Glaser L, Gordon JI: Purification and characterization of myristoyl CoA:protein N-myristoyltransferase, Proc Natl Acad Sci USA 84: 2708–2712, 1987
McIlhinney RAJ, Patel PB, McGlone K: Characterization of polyhistidine-tagged form of human myristoyl-CoA:protein N-myristoyltransferase produced in Escherichia coli. Eur J Biochem 222: 137–146, 1994
Peseckis SM, Resh MD: Fatty acyl transfer by human N-myristoyl transferase is dependent upon conserved cysteine and histidine residues. J Biol Chem 269: 30888–30892, 1994
Duronio RJ, Towler DA, Heuckeroth RO, Gordon JI: Disruption of the yeast N-myristoyl transferase gene caused recessive lethality. Science 243: 796–800, 1989
Lodge JK, Johnson RL, Weinberg RA, Gordon JI: Comparison of myristoyl CoA:protein N-myristoyltransferase from three pathogenic fungi: Cryptococcus neoformans, histoplasma capsulatum, and candida albicans. J Biol Chem 269: 2996–3009, 1994
Tubbs PK, Garland PB: Assay of coenzyme A and some acyl derivatives. Methods Enzymol 13: 535–551, 1968
Rudnick DA, McWherter CA, Rocque WJ, Lennon PJ, Getman DP, Gordon JI: Kinetic and structural evidence for a sequential ordered bi bi mechanism of catalysis of Saccharomyces cerevisiae myristoyl CoA:protein N-myristoyltransferase. J Biol Chem 266: 9732–9739, 1991
da Silva AM, Klein C: Rapid posttranslational myristoylation of a 68 kDa protein in D. discoideium. J Cell Biol 111: 401–407, 1990
Manenti S, Sorokine O, Van Dorsselaer A, Taniguchi H: Demyristoylation of the major substrate of protein kinase C (MARCKS) by the cytoplasmic fraction of brain synaptosomes. J Biol Chem 268: 6878–6881, 1993
McIlhinney RAJ, McGlone K: Evidence for a non-myristoylated pool of the 80 kDa protein kinase substrate of rat brain. Biochem J 271: 681–685, 1990
Manenti S, Sorokine O, Dorsselaer AV, Tanguchi H: Isolation of the non-myristoylated form of a major substrate of protein kinase-C (MARCKS) from bovine brain. J Biol Chem 269: 8309–8313, 1994
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Raju, R.V.S., Sharma, R.K. Coenzyme A dependent myristoylation and demyristoylation in the regulation of bovine spleen N-myristoyltransferase. Mol Cell Biochem 158, 107–113 (1996). https://doi.org/10.1007/BF00225835
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DOI: https://doi.org/10.1007/BF00225835