Selective peptidic and peptidomimetic inhibitors of Candida albicans myristoylCoA: Protein N-myristoyltransferase: A new approach to antifungal therapy

Sikorski, James A.; Devadas, Balekudru; Zupec, Mark E.; Freeman, Sandra K.; Brown, David; Lu, Hwang‐Fun; Nagarajan, Srinivasan; Mehta, Pramod P.; Wade, Arlene C.; Kishore, Nandini S.; Bryant, Martin L.; Getman, Daniel P.; McWherter, Charles A.; Gordon, Jeffrey I.

doi:10.1002/(sici)1097-0282(1997)43:1<43::aid-bip5>3.0.co;2-0

Cited by 50 publications

(33 citation statements)

References 51 publications

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“…For example, several studies have targeted the NMTs of Candida albicans and Cryptococcus neoformans, which are responsible for systemic fungal infections in immunocompromised humans, as treatment of such infections (43,(61)(62)(63)(64). NMTs have also been targeted for the development of antiviral drugs due to their essential role in viral replication.…”

Section: Discussionmentioning

confidence: 99%

Two N-Myristoyltransferase Isozymes Play Unique Roles in Protein Myristoylation, Proliferation, and Apoptosis

Ducker

Upson

French

et al. 2005

Molecular Cancer Research

103

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N-myristoyltransferases (NMT) add myristate to the NH 2 termini of certain proteins, thereby regulating their localization and/or biological function. Using RNA interference, this study functionally characterizes the two NMT isozymes in human cells. Unique small interfering RNAs (siRNA) for each isozyme were designed and shown to decrease NMT1 or NMT2 protein levels by at least 90%. Ablation of NMT1 inhibited cell replication associated with a loss of activation of c-Src and its target FAK as well as reduction of signaling through the c-Raf/mitogen-activated protein kinase/ extracellular signal-regulated kinase kinase/extracellular signal-regulated kinase pathway. Terminal deoxynucleotidyl transferase -mediated dUTP nick end labeling assays showed that depletion of either NMT isozyme induced apoptosis, with NMT2 having a 2.5-fold greater effect than NMT1. Western blot analyses revealed that loss of NMT2 shifted the expression of the BCL family of proteins toward apoptosis. Finally, intratumoral injection of siRNA for NMT1 or for both NMT1 and NMT2 inhibited tumor growth in vivo, whereas the same treatment with siRNA for NMT2 or negative control siRNA did not. Overall, the data indicate that NMT1 and NMT2 have only partially overlapping functions and that NMT1 is critical for tumor cell proliferation.

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Section: Discussionmentioning

confidence: 99%

Two N-Myristoyltransferase Isozymes Play Unique Roles in Protein Myristoylation, Proliferation, and Apoptosis

Ducker

Upson

French

et al. 2005

Molecular Cancer Research

103

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“…CELL gal drugs. This approach has met with success in C. albicans and C. neoformans, for which compounds that specifically inhibit myristoylation are being tested (11,12,26,29,42). To our knowledge, this option has not yet been explored for A. fumigatus, now the most common fungal pathogen found in autopsies of immunocompromised patients in European hospitals (19).…”

mentioning

confidence: 99%

Aspergillus nidulans swoF Encodes an N -Myristoyl Transferase

Shaw¹,

Momany

Momany³

2002

Eukaryot Cell

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Polar growth is a fundamental process in filamentous fungi and is necessary for disease initiation in many pathogenic systems. Previously, swoF was identified in Aspergillus nidulans as a single-locus, temperaturesensitive (ts) mutant aberrant in both polarity establishment and polarity maintenance. The swoF gene was cloned by complementation of the ts phenotype and sequenced. The derived protein sequence had high identity with N-myristoyl transferases (NMTs) found in fungi, plants, and animals. In addition, wild-type growth at restrictive temperature was partially restored by the addition of myristic acid to the growth medium. Sequencing revealed that the mutation in swoF changes the conserved aspartic acid 369 to a tyrosine. The predicted A. nidulans SwoF protein, SwoFp, was homology modeled based on crystal structures of NMTs from Saccharomyces cerevisiae and Candida albicans. The D369Y swoF mutation is on the opposite face of the protein, distal to the myristoyl coenzyme A and peptide substrate binding sites. In wild-type NMTs, D369 appears to stabilize a structural ␤-strand bend through two hydrogen bonds and an ionic interaction. These stabilizing bonds are abolished in the D369Y mutant. We hypothesize that a substrate of SwoFp must be myristoylated for proper polarity establishment and maintenance. The mutation prevents the proper function of SwoFp at restrictive temperature and thus blocks polar growth.

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“…As noted in the Introduction, the acyl-CoA substrate specificities of orthologous Nmts are highly conserved (30,52), whereas their peptide substrate specificities are somewhat different.…”

Section: Use Of Isogenic Strains Expressing Human or C Neoformans Nmmentioning

confidence: 99%

“…[25][26][27][28][29]; reviewed in Ref. 30). In addition, genetic tests have established that NMT is essential for the growth and viability of both C. neoformans and C. albicans (19,31).…”

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confidence: 99%

Genetic and Biochemical Studies Establish That the Fungicidal Effect of a Fully Depeptidized Inhibitor of Cryptococcus neoformans Myristoyl-CoA:ProteinN-Myristoyltransferase (Nmt) Is Nmt-dependent

Lodge¹,

Jackson-Machelski²,

Higgins³

et al. 1998

Journal of Biological Chemistry

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Cryptococcus neoformans is a fungal pathogen that causes chronic meningitis in 10% of patients with AIDS. Genetic and biochemical studies were conducted to determine whether myristoyl-CoA:protein N-myristoyltransferase (Nmt) is a target for development of a new class of fungicidal drugs. A single copy of a conditional lethal C. neoformans NMT allele was introduced into the fungal genome by homologous recombination. The allele (nmt487D) produces temperature-sensitive myristic acid auxotrophy. This phenotype is due, in part, to under-myristoylation of a cellular ADP ribosylation factor (Arf) and can be rescued by forced expression of human Nmt. Two isogenic strains with identical growth kinetics at 35°C were used to test the biological effects of an Nmt inhibitor. CPA8 contained a single copy of wild type C. neoformans NMT. HMC1 contained nmt487D plus 10 copies of human NMT. Since a single copy of nmt487D will not support growth at 35°C, survival of HMC1 depends upon its human Nmt. ALYASKLS-NH 2 , an inhibitor derived from an Arf, was fully depeptidized: p-[(2-methyl-1-imidazol-1-yl)butyl]phenyl-acetyl was used to represent the GLYA tetrapeptide, whereas SKLS was replaced with a chiral tyrosinol scaffold. Kinetic studies revealed K i (app) values of 1.8 ؎ 1 and 9 ؎ 2.4 M for purified fungal and human Nmts, respectively. The minimal inhibitory concentration of the compound was 2-fold lower for CPA8 compared with HMC1. A single dose of 100 M produced a 5-fold greater inhibition of protein synthesis in CPA8 versus HMC1. The strain specificity of these responses indicates that the fungicidal effect was Nmt-dependent. These two strains may be useful for screening chemical libraries for Nmt-based fungicidal compounds with relatively little activity against the human enzyme.

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Selective peptidic and peptidomimetic inhibitors of Candida albicans myristoylCoA: Protein N-myristoyltransferase: A new approach to antifungal therapy

Cited by 50 publications

References 51 publications

Two N-Myristoyltransferase Isozymes Play Unique Roles in Protein Myristoylation, Proliferation, and Apoptosis

Two N-Myristoyltransferase Isozymes Play Unique Roles in Protein Myristoylation, Proliferation, and Apoptosis

Aspergillus nidulans swoF Encodes an N -Myristoyl Transferase

Genetic and Biochemical Studies Establish That the Fungicidal Effect of a Fully Depeptidized Inhibitor of Cryptococcus neoformans Myristoyl-CoA:ProteinN-Myristoyltransferase (Nmt) Is Nmt-dependent

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