Validation and invalidation of chemical probes for the human N-myristoyltransferases
| Autor: | Kallemeijn, Wouter W, Lueg, Gregor A, Faronato, Monica, Hadavizadeh, Kate, Grocin, Andrea Goya, Ok-Ryul Song, Howell, Michael, Calado, Dinis P, Tate, Edward W |
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| Přispěvatelé: | The Royal Society, Cancer Research UK |
| Rok vydání: | 2019 |
| Předmět: |
Model organisms
metabolic tagging Biochemistry & Molecular Biology TRIS DIBENZYLIDENEACETONE DIPALLADIUM Immunology IMP-366 (DDD85646) PROTEIN D-NMAPPD (B13) 2-hydroxymyristic acid sortase A ligation Signalling & Oncogenes POTENTIAL-DRUG TARGET IMP-1088 chemical proteomics Chemical Biology & High Throughput Science & Technology MYRISTOYL-COA DBA PALLADIUM Stem Cells FOS: Clinical medicine Tumour Biology CANCER ESSENTIAL ENZYME APOPTOSIS N-myristoylation DISCOVERY Tris-DBA palladium N-myristoyltransferases (NMT) INHIBITORS Genetics & Genomics Life Sciences & Biomedicine Developmental Biology |
| Popis: | On-target, cell-active chemical probes are of fundamental importance in both chemical and cell biology, whereas the application of poorly-characterised probes often leads to invalid conclusions.Human N-myristoyltransferase (NMT) has attracted increasing interest as a 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 which used off-target compounds. |
| Databáze: | OpenAIRE |
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