Validation and invalidation of chemical probes for the human N-myristoyltransferases.
Wouter W Kallemeijn
Gregor A Lueg
Monica Faronato
Kate Hadavizadeh
Andrea Goya Grocin
Ok-Ryul Song
Michael Howell
Dinis P Calado
Edward W Tate
10779/crick.11522910.v1
https://crick.figshare.com/articles/journal_contribution/Validation_and_invalidation_of_chemical_probes_for_the_human_N-myristoyltransferases_/11522910
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.
2020-01-06 17:42:54
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
Calado FC001057
HTS
CS-ack
Tate - sat