2. Academic Validation
  3. Validation and Invalidation of Chemical Probes for the Human N-myristoyltransferases

Validation and Invalidation of Chemical Probes for the Human N-myristoyltransferases

  • Cell Chem Biol. 2019 Jun 20;26(6):892-900.e4. doi: 10.1016/j.chembiol.2019.03.006.
Wouter W Kallemeijn 1,  Gregor A Lueg 2,  Monica Faronato 2,  Kate Hadavizadeh 1,  Andrea Goya Grocin 1,  Ok-Ryul Song 3,  Michael Howell 3,  Dinis P Calado 4,  Edward W Tate 5
Affiliations

Affiliations

  • 1 Department of Chemistry, Imperial College London, Molecular Research Science Hub, 80 Wood Lane, London W12 0BZ, UK.
  • 2 Department of Chemistry, Imperial College London, Molecular Research Science Hub, 80 Wood Lane, London W12 0BZ, UK; The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
  • 3 The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
  • 4 The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK; Peter Gorer Department of Immunobiology, School of Immunology & Microbial Sciences, King's College London, London SE1 9RT, UK.
  • 5 Department of Chemistry, Imperial College London, Molecular Research Science Hub, 80 Wood Lane, London W12 0BZ, UK; The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK. Electronic address: [email protected].
PMID: 31006618 DOI: 10.1016/j.chembiol.2019.03.006
Abstract

On-target, cell-active chemical probes are of fundamental importance in chemical and Cell Biology, whereas poorly characterized probes often lead to invalid conclusions. Human N-myristoyltransferase (NMT) has attracted increasing interest as target in Cancer and infectious diseases. Here we report an in-depth comparison of five compounds widely applied as human NMT inhibitors, using a combination of quantitative whole-proteome N-myristoylation profiling, biochemical Enzyme assays, cytotoxicity, in-cell protein synthesis, and cell-cycle assays. We find that N-myristoylation is unaffected by 2-hydroxymyristic acid (100 μM), D-NMAPPD (30 μM), or Tris-DBA palladium (10 μM), with the latter compounds causing cytotoxicity through mechanisms unrelated to NMT. In contrast, drug-like inhibitors IMP-366 (DDD85646) and IMP-1088 delivered complete and specific inhibition of N-myristoylation in a range of cell lines at 1 μM and 100 nM, respectively. This study enables the selection of appropriate on-target probes for future studies and suggests the need for reassessment of previous studies that used off-target compounds.

Keywords

2-hydroxymyristic acid; D-NMAPPD (B13); IMP-1088; IMP-366 (DDD85646); N-myristoylation; N-myristoyltransferases (NMT); Tris-DBA palladium; chemical proteomics; metabolic tagging; sortase A ligation.

Figures
Products