Mechanism of subunit interaction at ketosynthase-dehydratase junctions in trans-AT polyketide synthases

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...

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Main Authors: Jenner, Matthew, Kosol, Simone, Griffiths, Daniel, Prasongpholchai, Panward, Manzi, Lucio, Barrow, Andrew S., Moses, John E., Oldham, Neil J., Lewandowski, Jozef, Challis, Gregory
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Published: Nature Publishing Group 2018
Online Access:https://eprints.nottingham.ac.uk/49829/
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author Jenner, Matthew
Kosol, Simone
Griffiths, Daniel
Prasongpholchai, Panward
Manzi, Lucio
Barrow, Andrew S.
Moses, John E.
Oldham, Neil J.
Lewandowski, Jozef
Challis, Gregory
author_facet Jenner, Matthew
Kosol, Simone
Griffiths, Daniel
Prasongpholchai, Panward
Manzi, Lucio
Barrow, Andrew S.
Moses, John E.
Oldham, Neil J.
Lewandowski, Jozef
Challis, Gregory
author_sort Jenner, Matthew
building Nottingham Research Data Repository
collection Online Access
description 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.
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spelling nottingham-498292020-05-04T19:26:06Z https://eprints.nottingham.ac.uk/49829/ Mechanism of subunit interaction at ketosynthase-dehydratase junctions in trans-AT 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 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. Nature Publishing Group 2018-01-08 Article PeerReviewed Jenner, Matthew, Kosol, Simone, Griffiths, Daniel, Prasongpholchai, Panward, Manzi, Lucio, Barrow, Andrew S., Moses, John E., Oldham, Neil J., Lewandowski, Jozef and Challis, Gregory (2018) Mechanism of subunit interaction at ketosynthase-dehydratase junctions in trans-AT polyketide synthases. Nature Chemical Biology, 14 . pp. 270-275. ISSN 1552-4450 https://www.nature.com/articles/nchembio.2549 doi:10.1038/nchembio.2549 doi:10.1038/nchembio.2549
spellingShingle Jenner, Matthew
Kosol, Simone
Griffiths, Daniel
Prasongpholchai, Panward
Manzi, Lucio
Barrow, Andrew S.
Moses, John E.
Oldham, Neil J.
Lewandowski, Jozef
Challis, Gregory
Mechanism of subunit interaction at ketosynthase-dehydratase junctions in trans-AT polyketide synthases
title Mechanism of subunit interaction at ketosynthase-dehydratase junctions in trans-AT polyketide synthases
title_full Mechanism of subunit interaction at ketosynthase-dehydratase junctions in trans-AT polyketide synthases
title_fullStr Mechanism of subunit interaction at ketosynthase-dehydratase junctions in trans-AT polyketide synthases
title_full_unstemmed Mechanism of subunit interaction at ketosynthase-dehydratase junctions in trans-AT polyketide synthases
title_short Mechanism of subunit interaction at ketosynthase-dehydratase junctions in trans-AT polyketide synthases
title_sort mechanism of subunit interaction at ketosynthase-dehydratase junctions in trans-at polyketide synthases
url https://eprints.nottingham.ac.uk/49829/
https://eprints.nottingham.ac.uk/49829/
https://eprints.nottingham.ac.uk/49829/