Mechanism of intersubunit ketosynthase–dehydratase interaction in polyketide synthases

Jenner, Matthew; Kosol, Simone; Griffiths, Daniel; Prasongpholchai, Panward; Manzi, Lucio; Barrow, Andrew S.; Moses, John E.; Oldham, Neil J.; Lewandowski, Jozef; Challis, Gregory

Abstract

Modular polyketide synthases (PKSs) produce numerous structurally complex natural products with diverse applications in medicine and agriculture. They typically consist of several multienzyme subunits that utilize structurally-defined docking domains (DDs) at their N- and C-termini to ensure correct assembly into functional multi-protein complexes. Here we report a fundamentally different mechanism for subunit assembly in trans-AT modular PKSs at the junction between ketosynthase (KS) and dehydratase (DH) domains. This involves direct interaction of a largely unstructured docking domain (DD) at the C-terminus of the KS with the surface of the downstream DH. Acyl transfer assays and mechanism-based cross-linking established that the DD is required for the KS to communicate with the acyl carrier protein appended to the DH. Two distinct regions for binding of the DD to the DH were identified using NMR spectroscopy, carbene foot-printing and mutagenesis, providing a foundation for future elucidation of the molecular basis for interaction specificity.

Citation

Jenner, M., Kosol, S., Griffiths, D., Prasongpholchai, P., Manzi, L., Barrow, A. S., …Challis, G. (2018). Mechanism of intersubunit ketosynthase–dehydratase interaction in polyketide synthases. Nature Chemical Biology, 14, 270-275. https://doi.org/10.1038/nchembio.2549

• Journal Article Type: Article
• Acceptance Date: Nov 15, 2017
• Online Publication Date: Jan 8, 2018
• Publication Date: 2018-03
• Deposit Date: Feb 16, 2018
• Publicly Available Date: Jul 9, 2018
• Journal: Nature Chemical Biology
• Print ISSN: 1552-4450
• Electronic ISSN: 1552-4469
• Publisher: Nature Publishing Group
• Peer Reviewed: Peer Reviewed
• Volume: 14
• Pages: 270-275
• DOI: https://doi.org/10.1038/nchembio.2549
• Public URL: https://nottingham-repository.worktribe.com/output/903708
• Publisher URL: https://www.nature.com/articles/nchembio.2549
• Contract Date: Feb 16, 2018