A conserved motif flags acyl carrier proteins for β-branching in polyketide synthesis

Anthony S. Haines*, Xu Dong, Zhongshu Song, Rohit Farmer, Christopher Williams, Joanne Hothersall, Eliza Ploskon, Pakorn Wattana-amorn, Elton R. Stephens, Erika Yamada, Rachel Gurney, Yuiko Takebayashi, Joleen Masschelein, Russell J. Cox, Rob Lavigne, Christine L. Willis, Thomas J. Simpson, John Crosby, Peter J. Winn, Christopher M. ThomasMatthew P. Crump

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)

33 Citations (Scopus)
556 Downloads (Pure)

Abstract

Type I polyketide synthases often use programmed β-branching, via enzymes of a 'hydroxymethylglutaryl-CoA synthase (HCS) cassette', to incorporate various side chains at the second carbon from the terminal carboxylic acid of growing polyketide backbones. We identified a strong sequence motif in acyl carrier proteins (ACPs) where β-branching is known to occur. Substituting ACPs confirmed a correlation of ACP type with β-branching specificity. Although these ACPs often occur in tandem, NMR analysis of tandem β-branching ACPs indicated no ACP-ACP synergistic effects and revealed that the conserved sequence motif forms an internal core rather than an exposed patch. Modeling and mutagenesis identified ACP helix III as a probable anchor point of the ACP-HCS complex whose position is determined by the core. Mutating the core affects ACP functionality, whereas ACP-HCS interface substitutions modulate system specificity. Our method for predicting β-carbon branching expands the potential for engineering new polyketides and lays a basis for determining specificity rules.
Original languageEnglish
Pages (from-to)685-692
Number of pages8
JournalNature Chemical Biology
Volume9
Issue number11
Early online date22 Sep 2013
DOIs
Publication statusPublished - Nov 2013

Structured keywords

  • Bristol BioDesign Institute

Keywords

  • PSEUDOMONAS-FLUORESCENS NCIMB-10586
  • CYANOBACTERIUM LYNGBYA-MAJUSCULA
  • BIOSYNTHESIS GENE-CLUSTER
  • NMR-SPECTROSCOPY
  • NATURAL-PRODUCT
  • ANTIBIOTIC MUPIROCIN
  • BACILLUS-SUBTILIS
  • DIPOLAR COUPLINGS
  • COELICOLOR A3(2)
  • CURACIN-A

Fingerprint Dive into the research topics of 'A conserved motif flags acyl carrier proteins for β-branching in polyketide synthesis'. Together they form a unique fingerprint.

  • Projects

    Cite this

    Haines, A. S., Dong, X., Song, Z., Farmer, R., Williams, C., Hothersall, J., Ploskon, E., Wattana-amorn, P., Stephens, E. R., Yamada, E., Gurney, R., Takebayashi, Y., Masschelein, J., Cox, R. J., Lavigne, R., Willis, C. L., Simpson, T. J., Crosby, J., Winn, P. J., ... Crump, M. P. (2013). A conserved motif flags acyl carrier proteins for β-branching in polyketide synthesis. Nature Chemical Biology, 9(11), 685-692. https://doi.org/10.1038/nchembio.1342