Identification of Protein Palmitoylation Inhibitors from a Scaffold Ranking Library.

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  • Additional Information
    • Source:
      Publisher: Bentham Science Publishers Country of Publication: United Arab Emirates NLM ID: 9810948 Publication Model: Print Cited Medium: Internet ISSN: 1875-5402 (Electronic) Linking ISSN: 13862073 NLM ISO Abbreviation: Comb Chem High Throughput Screen Subsets: MEDLINE
    • Publication Information:
      Publication: Saif Zone, Sharjah, U.A.E. : Bentham Science Publishers
      Original Publication: Hilversum, Netherlands ; Miami, FL : Bentham Science Publishers, c1998-
    • Subject Terms:
      Acyltransferases/*antagonists & inhibitors ; High-Throughput Screening Assays/*methods ; Lipoylation/*drug effects ; Membrane Proteins/*antagonists & inhibitors ; Protein Processing, Post-Translational/*drug effects ; Saccharomyces cerevisiae Proteins/*antagonists & inhibitors; Animals ; Humans ; Palmitic Acid/chemistry ; Piperazine ; Piperazines ; Saccharomyces cerevisiae ; Small Molecule Libraries/chemistry
    • Abstract:
      The addition of palmitoyl moieties to proteins regulates their membrane targeting, subcellular localization, and stability. Dysregulation of the enzymes which catalyzed the palmitoyl addition and/or the substrates of these enzymes have been linked to cancer, cardiovascular, and neurological disorders, implying these enzymes and substrates are valid targets for pharmaceutical intervention. However, current chemical modulators of zDHHC PAT enzymes lack specificity and affinity, underscoring the need for screening campaigns to identify new specific, high affinity modulators. This report describes a mixture based screening approach to identify inhibitors of Erf2 activity. Erf2 is the Saccharomyces cerevisiae PAT responsible for catalyzing the palmitoylation of Ras2, an ortholog of the human Ras oncogene proteins. A chemical library developed by the Torrey Pines Institute for Molecular Studies consists of more than 30 million compounds designed around 68 molecular scaffolds that are systematically arranged into positional scanning and scaffold ranking formats. We have used this approach to identify and characterize several scaffold backbones and R-groups that reduce or eliminate the activity of Erf2 in vitro. Here, we present the analysis of one of the scaffold backbones, bis-cyclic piperazine. We identified compounds that inhibited Erf2 auto-palmitoylation activity using a fluorescence-based, coupled assay in a high throughput screening (HTS) format and validated the hits utilizing an orthogonal gel-based assay. Finally, we examined the effects of the compounds on cell growth in a yeast cell-based assay. Based on our results, we have identified specific, high affinity palmitoyl transferase inhibitors that will serve as a foundation for future compound design.
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    • Grant Information:
      R21 NS090160 United States NS NINDS NIH HHS
    • Accession Number:
      0 (Membrane Proteins)
      0 (Piperazines)
      0 (Saccharomyces cerevisiae Proteins)
      0 (Small Molecule Libraries)
      1RTM4PAL0V (Piperazine)
      2V16EO95H1 (Palmitic Acid)
      EC 2.3.- (Acyltransferases)
      EC 2.3.1.- (ERF2 protein, S cerevisiae)
    • Publication Date:
      Date Created: 20160325 Date Completed: 20170728 Latest Revision: 20190923
    • Publication Date:
      20240829
    • Accession Number:
      PMC5068503
    • Accession Number:
      10.2174/1386207319666160324123844
    • Accession Number:
      27009891