Heterologous Expression, Biosynthetic Studies, and Ecological Function of the Selective Gq-Signaling Inhibitor FR900359

Angew Chem Int Ed Engl. 2018 Jan 15;57(3):836-840. doi: 10.1002/anie.201707996. Epub 2017 Dec 20.

Abstract

The cyclic depsipeptide FR900359 (FR), isolated from the tropical plant Ardisia crenata, is a strong and selective inhibitor of Gq proteins, making it an indispensable pharmacological tool to study Gq-related processes, as well as a promising drug candidate. Gq inhibition is a novel mode of action for defense chemicals and crucial for the ecological function of FR, as shown by in vivo experiments in mice, its affinity to insect Gq proteins, and insect toxicity studies. The uncultured endosymbiont of A. crenata was sequenced, revealing the FR nonribosomal peptide synthetase (frs) gene cluster. We here provide a detailed model of FR biosynthesis, supported by in vitro enzymatic and bioinformatic studies, and the novel analogue AC-1, which demonstrates the flexibility of the FR starter condensation domains. Finally, expression of the frs genes in E. coli led to heterologous FR production in a cultivable, bacterial host for the first time.

Keywords: FR900359; G proteins; biosynthesis; heterologous expression; natural products.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bombyx / metabolism
  • Chromosomes, Artificial, Bacterial
  • Computational Biology
  • Depsipeptides / biosynthesis*
  • Depsipeptides / metabolism
  • Depsipeptides / pharmacology*
  • Escherichia coli / genetics
  • GTP-Binding Protein alpha Subunits, Gq-G11 / metabolism*
  • Gene Transfer Techniques
  • HEK293 Cells
  • Humans
  • Insect Proteins / metabolism*
  • Multigene Family
  • Peptide Synthases / genetics
  • Primulaceae / chemistry
  • Sf9 Cells
  • Signal Transduction / drug effects*
  • Tandem Mass Spectrometry

Substances

  • Depsipeptides
  • FR900359
  • Insect Proteins
  • GTP-Binding Protein alpha Subunits, Gq-G11
  • Peptide Synthases
  • non-ribosomal peptide synthase