Affinage · Plasmodium

NMT

PF3D7_1412800 · glycylpeptide N-tetradecanoyltransferase

Annotated
2026-09-08
33 papers in source corpus 11 papers cited in narrative 11 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

PF3D7_1412800 (NMT) is a single-copy, essential glycylpeptide N-tetradecanoyltransferase of Plasmodium that transfers myristate from myristoyl-CoA to the N-terminal glycine of substrate proteins during asexual blood-stage development (PMID:10816442, PMID:23245327). The recombinant enzyme is catalytically active on synthetic peptide substrates, its catalytic and substrate-binding residues are conserved, and it displays a chemical-inhibitor sensitivity distinct from the human enzyme (PMID:10816442). NMT myristoylates a defined set of parasite substrates including adenylate kinase 2 (AK2), with which it also forms a heterodimeric complex in vitro (PMID:18973776), and the glideosome protein GAP45, which is dually acylated by N-terminal myristoylation and palmitoylation to support membrane association (PMID:16750579). Enzyme activity is regulated by the acyl-CoA binding protein ACBD6, which protects NMT from lauryl-CoA inhibition and enforces preferential use of myristoyl-CoA over competing palmitoyl-CoA (PMID:30642881). Pharmacological inhibition disrupts parasite development at three stages—early schizogony, late schizogony/egress, and erythrocyte invasion—by reducing myristoylation and destabilizing or mislocalizing substrate proteins, with substrate-specific consequences: for GAP45, loss of myristoylation spares localization and glideosome assembly yet is still required for invasion (PMID:34695132, PMID:29363940). High-resolution ternary crystal structures of Plasmodium NMTs capture the enzyme with myristoyl-CoA and peptidomimetic or drug-like inhibitors bound in the peptide-substrate pocket, and a single substitution in that pocket confers resistance to one inhibitor series, validating the active site as an antimalarial target (PMID:31080074, PMID:22439843, PMID:25230674, PMID:39655507).

Mechanistic history

Synthesis pass · year-by-year structured walk · 10 steps
  1. 2000 High

    Establishing that Plasmodium encodes a genuine myristoyltransferase answered whether the parasite possesses this enzymatic machinery and whether its catalytic properties differ from the human host enzyme.

    Evidence Cloning, sequence comparison, recombinant expression in E. coli, in vitro activity assay on a synthetic peptide, and chemical-modification inhibition comparisons

    PMID:10816442

    Open questions at the time
    • Endogenous substrate repertoire not defined
    • Subcellular localization in the parasite not established
    • Essentiality not yet tested genetically
  2. 2006 Medium

    Identifying GAP45 as a dually acylated NMT substrate connected the enzyme to a specific membrane-targeting event in merozoites.

    Evidence Metabolic labeling with [3H]-myristate and [3H]-palmitate, immunoprecipitation, and mass spectrometry in P. falciparum

    PMID:16750579

    Open questions at the time
    • Functional requirement of GAP45 myristoylation for invasion not yet dissected here
    • Interplay of myristoylation and palmitoylation timing unresolved
  3. 2008 High

    Demonstrating myristoylation of intact AK2 and a stable NMT–AK2 heterodimer showed NMT acts on a full-length parasite protein and can form a defined complex with a substrate.

    Evidence Co-expression in E. coli with myristate, purification, mass spectrometry, and in vitro protein-protein interaction

    PMID:18973776

    Open questions at the time
    • Physiological relevance of the heterodimer in vivo unknown
    • Whether heterodimerization reflects a general substrate-engagement mode or is AK2-specific is unclear
  4. 2012 High

    Conditional knockdown established genetically that NMT is essential for blood-stage survival, moving it from a biochemically active enzyme to a validated dependency.

    Evidence Tetracycline-repressible transactivator knockdown and viability assay in P. berghei

    PMID:23245327

    Open questions at the time
    • Species is P. berghei, not P. falciparum
    • Which substrate(s) drive the lethal phenotype not identified
    • Stage-specific requirements not resolved
  5. 2012 High

    The first Plasmodium NMT co-crystal structure with a screening hit defined the inhibitor binding mode and initiated structure-guided antimalarial design.

    Evidence High-throughput screening, high-resolution X-ray co-crystal structure, and structure-activity relationship analysis of P. vivax NMT

    PMID:22439843

    Open questions at the time
    • Structure is from P. vivax NMT
    • Cellular efficacy not addressed by structure alone
  6. 2014 High

    A ternary NMT–CoA/myristoylated-peptide product structure provided direct mechanistic insight into the catalytic reaction and comparative binding across parasite NMTs.

    Evidence X-ray crystallography of ternary product complexes with P. falciparum and Leishmania NMTs plus enzyme inhibition assays

    PMID:25230674

    Open questions at the time
    • Catalytic mechanism inferred from static structures
    • Substrate-selectivity determinants not fully mapped
  7. 2019 Medium

    Discovery that ACBD6 protects NMT from lauryl-CoA inhibition and enforces myristoyl-CoA preference identified a regulatory layer controlling acyl-CoA specificity under competing-substrate conditions.

    Evidence In vitro enzymatic activity and acyl-CoA competition assays with recombinant P. falciparum proteins

    PMID:30642881

    Open questions at the time
    • Regulatory interaction shown in vitro; in vivo role in the parasite not established
    • Whether ACBD6 physically docks onto NMT not resolved
  8. 2019 High

    Resistance mapping to a single substrate-pocket substitution confirmed that NMT itself is the antimalarial target of a specific inhibitor series and revealed a chemically distinct series that circumvents it.

    Evidence In vitro resistance selection, crystallography, enzyme activity, thermal stability, and surface plasmon resonance

    PMID:31080074

    Open questions at the time
    • Fitness cost of the resistance mutation not characterized
    • Clinical relevance of resistance not established
  9. 2021 High

    Chemical inhibition combined with substrate glycine-to-alanine mutagenesis dissected the pleiotropic, stage-specific consequences of NMT loss and showed substrate-specific myristoylation outcomes.

    Evidence NMT inhibitor treatment, N-terminal glycine-to-alanine substitution, chemical proteomics, microscopy, and egress/invasion assays in P. falciparum

    PMID:34695132

    Open questions at the time
    • Mechanism by which unmyristoylated GAP45 fails at invasion despite normal localization is unresolved
    • Only a subset of the 16 affected substrates were functionally validated
  10. 2025 High

    A 1.8 Å ternary structure with IMP-1088 showed the drug engages the parasite peptide-binding pocket much as it does the human enzyme, informing selectivity considerations.

    Evidence 1.8 Å X-ray ternary complex of P. vivax NMT with myristoyl-CoA and IMP-1088

    PMID:39655507

    Open questions at the time
    • Structure from P. vivax NMT
    • Selectivity margin over human NMT not quantified from structure alone

Open questions

Synthesis pass · forward-looking unresolved questions
  • How NMT selects among its diverse substrates, how ACBD6 regulation operates in the living parasite, and how myristoylation controls each stage-specific function at the molecular level remain open.
  • Full endogenous substrate set and selectivity code undefined
  • In vivo role of ACBD6–NMT regulation unconfirmed
  • Mechanistic basis of GAP45's myristoylation-dependent invasion function unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification
Partners
ACBD6 — ACYL-COA BINDING PROTEIN THAT REGULATES NMT ACYL-COA SPECIFICITY [#3]ADENYLATE KINASE 2 (AK2) — SUBSTRATE AND HETERODIMER PARTNER [#1]GAP45 — DUALLY ACYLATED SUBSTRATE [#10, #4]
Complex memberships
NMT–AK2 heterodimer (in vitro) [#1]

Evidence

Reading pass · 11 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2000 P. falciparum NMT (PF3D7_1412800) was cloned from P. falciparum, shown to be single-copy with mRNA expressed in asexual blood stages, and the recombinant protein expressed in E. coli was demonstrated to have myristoyltransferase enzymatic activity using a synthetic peptide substrate. The active-site residues important for substrate binding and catalysis in other species are conserved. Unlike human NMT, P. falciparum NMT activity was not inhibited by diethylpyrocarbonate or iodoacetamide, indicating a different pattern of inhibitor sensitivity by chemical modification. Gene cloning, cDNA/genomic sequence comparison, recombinant protein expression in E. coli, in vitro enzymatic activity assay with synthetic peptide substrate, chemical modification inhibition assays The Biochemical journal High 10816442
2008 P. falciparum NMT (PF3D7_1412800) was co-expressed with adenylate kinase 2 (AK2) in E. coli in the presence of myristate, and mass spectrometry confirmed that NMT catalyzes myristoylation of the intact malarial AK2 protein. Furthermore, AK2 and NMT were shown to interact strongly with each other, forming a heterodimeric protein complex in vitro. Heterologous co-expression in E. coli, protein purification, mass spectrometry for myristoylation confirmation, in vitro protein-protein interaction (heterodimer formation) Molecular and biochemical parasitology High 18973776
2012 Tetracycline-induced conditional knockdown of N-myristoyltransferase (NMT) in P. berghei using a TetR-based transactivator system resulted in a dramatic reduction in parasite viability, establishing that NMT is essential for P. berghei blood-stage parasite survival. Conditional knockdown using tetracycline-repressible transactivator (TetR) system, parasite viability assay Cell host & microbe High 23245327
2019 ACBD6 (an acyl-CoA binding domain protein) supports and regulates the activity of P. falciparum NMT (PF3D7_1412800). ACBD6 protects NMT from inhibition by lauryl-CoA and promotes preferential use of myristoyl-CoA substrate, thereby enhancing N-myristoylation under substrate-limiting conditions where palmitoyl-CoA competes with myristoyl-CoA for NMT binding. In vitro enzymatic activity assays with recombinant proteins, acyl-CoA competition assays Journal of lipid research Medium 30642881
2021 Treatment of P. falciparum with NMT inhibitor IMP-1002 during schizogony blocked parasite egress from erythrocytes, disrupted rhoptry protein localization, and impaired erythrocyte invasion. Sixteen NMT substrate proteins showed significantly reduced myristoylation upon NMTi treatment, of which six were substantially reduced in abundance. Replacement of the N-terminal glycine with alanine (preventing myristoylation) in four of these substrates substantially impaired parasite fitness. For GAP45 specifically, loss of myristoylation did not affect protein localization or glideosome assembly but was essential for erythrocyte invasion, demonstrating mechanistically distinct consequences of myristoylation for individual substrates. NMT inhibition thus disrupts parasite development at three stages: early schizogony (pseudoschizont formation), late schizogony (rhoptry formation/egress), and invasion. Chemical biology (NMT inhibitor treatment), genetic modification (N-terminal glycine-to-alanine substitution), parasite viability screen, chemical proteomics for myristoylation profiling, fluorescence microscopy for protein localization, parasite egress and invasion assays PLoS biology High 34695132
2019 In vitro resistance to a specific NMT inhibitor series was mediated by a single amino acid substitution in the NMT substrate-binding pocket, validated by crystallography, enzyme activity assays, stability assays, and surface plasmon resonance. A second inhibitor series was identified that was unaffected by this substitution. In vitro resistance selection, crystallography, enzyme activity assays, thermal stability assays, surface plasmon resonance Cell chemical biology High 31080074
2012 Crystal structure of P. vivax NMT in complex with a hit compound from high-throughput screening was obtained at high resolution, revealing the novel binding mode of the inhibitor in the NMT active site. Structure-activity relationships defined chemical groups relevant for activity and selectivity. High-throughput screening, X-ray crystallography (high-resolution co-crystal structure with inhibitor), structure-activity relationship analysis Journal of medicinal chemistry High 22439843
2014 High-resolution crystal structures of peptidomimetic inhibitors in complex with P. falciparum NMT and Leishmania NMT were obtained, including the first crystal structure evidence for a ternary NMT–Coenzyme A/myristoylated peptide product complex, revealing comparative binding modes between Plasmodium and Leishmania NMTs. X-ray crystallography (ternary complex structure), enzyme inhibition assays Organic & biomolecular chemistry High 25230674
2018 NMT inhibition in P. falciparum results in incorrect localization of myristoylated substrate proteins, disrupting their membrane interactions and causing pleiotropic effects on parasite development, growth, and multiplication. NMT substrates include proteins involved in secretory transport and apicomplexan-specific processes including gliding motility. Review/perspective synthesizing chemical biology and inhibitor studies; substrate localization experiments cited from prior work ACS infectious diseases Medium 29363940
2025 A 1.8 Å resolution ternary crystal structure of P. vivax NMT in complex with myristoyl-CoA and the inhibitor IMP-1088 was determined, showing that IMP-1088 binds PvNMT similarly to how it binds human NMT, occupying the peptide substrate-binding pocket. X-ray crystallography (1.8 Å ternary complex structure) Acta crystallographica. Section F, Structural biology communications High 39655507
2006 GAP45 in P. falciparum merozoites undergoes dual acylation: N-terminal myristoylation (catalyzed by NMT) and palmitoylation. This dual acylation is important for membrane association of GAP45. Metabolic labeling with [3H]-myristate and [3H]-palmitate, immunoprecipitation, mass spectrometry Molecular and biochemical parasitology Medium 16750579

Source papers

Stage 0 corpus · 33 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2006 Dual acylation of the 45 kDa gliding-associated protein (GAP45) in Plasmodium falciparum merozoites. Molecular and biochemical parasitology 90 16750579
2012 Selective inhibitors of protozoan protein N-myristoyltransferases as starting points for tropical disease medicinal chemistry programs. PLoS neglected tropical diseases 78 22545171
2012 A tetracycline-repressible transactivator system to study essential genes in malaria parasites. Cell host & microbe 76 23245327
2014 Design and synthesis of high affinity inhibitors of Plasmodium falciparum and Plasmodium vivax N-myristoyltransferases directed by ligand efficiency dependent lipophilicity (LELP). Journal of medicinal chemistry 59 24641010
2012 Discovery of Plasmodium vivax N-myristoyltransferase inhibitors: screening, synthesis, and structural characterization of their binding mode. Journal of medicinal chemistry 58 22439843
2010 Identification of inhibitors for putative malaria drug targets among novel antimalarial compounds. Molecular and biochemical parasitology 58 20813141
2012 Design and synthesis of inhibitors of Plasmodium falciparum N-myristoyltransferase, a promising target for antimalarial drug discovery. Journal of medicinal chemistry 51 23035716
2000 Characterization of N-myristoyltransferase from Plasmodium falciparum. The Biochemical journal 48 10816442
2018 N-Myristoylation as a Drug Target in Malaria: Exploring the Role of N-Myristoyltransferase Substrates in the Inhibitor Mode of Action. ACS infectious diseases 46 29363940
2012 Discovery of novel and ligand-efficient inhibitors of Plasmodium falciparum and Plasmodium vivax N-myristoyltransferase. Journal of medicinal chemistry 46 23170970
2019 Structure-Guided Identification of Resistance Breaking Antimalarial N‑Myristoyltransferase Inhibitors. Cell chemical biology 33 31080074
2023 Identification of potent and selective N-myristoyltransferase inhibitors of Plasmodium vivax liver stage hypnozoites and schizonts. Nature communications 31 37669940
2017 New developments in probing and targeting protein acylation in malaria, leishmaniasis and African sleeping sickness. Parasitology 28 28270257
2014 Peptidomimetic inhibitors of N-myristoyltransferase from human malaria and leishmaniasis parasites. Organic & biomolecular chemistry 27 25230674
2020 Identification of Selective Inhibitors of Plasmodium N-Myristoyltransferase by High-Throughput Screening. Journal of medicinal chemistry 23 31850752
2021 Inhibition of protein N-myristoylation blocks Plasmodium falciparum intraerythrocytic development, egress and invasion. PLoS biology 18 34695132
2019 ACBD6 protein controls acyl chain availability and specificity of the N-myristoylation modification of proteins. Journal of lipid research 17 30642881
2016 Structure-guided optimization of quinoline inhibitors of Plasmodium N-myristoyltransferase. MedChemComm 17 28626547
2015 Discovery of pyridyl-based inhibitors of Plasmodium falciparum N-myristoyltransferase. MedChemComm 15 26962430
2009 Homology modeling and molecular dynamics simulation of N-myristoyltransferase from protozoan parasites: active site characterization and insights into rational inhibitor design. Journal of computer-aided molecular design 13 19370313
2008 Myristoylated adenylate kinase-2 of Plasmodium falciparum forms a heterodimer with myristoyltransferase. Molecular and biochemical parasitology 13 18973776
2020 QSAR studies on benzothiophene derivatives as Plasmodium falciparum N-myristoyltransferase inhibitors: Molecular insights into affinity and selectivity. Drug development research 9 32045013
2015 Homology modeling and molecular dynamics simulation of N-myristoyltransferase from Plasmodium falciparum: an insight into novel antimalarial drug design. Journal of molecular modeling 7 25663521
2022 Identification of potential inhibitors for N-myristoyltransferase (NMT) protein of Plasmodium vivax. Journal of biomolecular structure & dynamics 6 36002266
2018 N-myristoyltransferases inhibitory activity of ellagitannins from Terminalia bentzoë (L.) L. f. subsp. bentzoë. Fitoterapia 6 30342177
2025 Structure of Plasmodium vivaxN-myristoyltransferase with inhibitor IMP-1088: exploring an NMT inhibitor for antimalarial therapy. Acta crystallographica. Section F, Structural biology communications 5 39655507
2012 Analyzing a Potential Drug Target N-Myristoyltransferase of Plasmodium falciparum Through In Silico Approaches. Journal of global infectious diseases 5 22529627
2024 Current trends to design antimalarial drugs targeting N-myristoyltransferase. Future microbiology 4 39440556
2024 Exploring Subsite Selectivity within Plasmodium vivax N-Myristoyltransferase Using Pyrazole-Derived Inhibitors. Journal of medicinal chemistry 2 38680035
2024 Selective Bias Virtual Screening for Discovery of Promising Antimalarial Candidates targeting Plasmodium N-Myristoyltransferase. Research square 1 38463971
2026 Design, Synthesis, and Antimalarial Evaluation of Novel Quinazolin-4(3H)‑one Derivatives with Molecular Modeling Insights into Target Selectivity. ACS omega 0 42368058
2025 Molecular docking analysis of N-myristoyl-transferase with small molecules from the ZINC database for screening potential anti-malarial drugs. Bioinformation 0 40978577
2025 N-Myristoyltransferase Inhibition in Parasitic Pathogens: Insights from Computer-Aided Drug Design. Molecules (Basel, Switzerland) 0 41011596