Towards a comprehensive understanding of the structural dynamics of a bacterial diterpene synthase during catalysis

Ronja Driller, Sophie Janke, Monika Fuchs, Evelyn Warner, Anil R. Mhashal, Dan Thomas Major, Mathias Christmann, Thomas Brück, Bernhard Loll

Research output: Contribution to journalArticlepeer-review

53 Scopus citations

Abstract

Terpenes constitute the largest and structurally most diverse natural product family. Most terpenoids exhibit a stereochemically complex macrocyclic core, which is generated by C–C bond forming of aliphatic oligo-prenyl precursors. This reaction is catalysed by terpene synthases (TPSs), which are capable of chaperoning highly reactive carbocation intermediates through an enzyme-specific reaction. Due to the instability of carbocation intermediates, the proteins’ structural dynamics and enzyme:substrate interactions during TPS catalysis remain elusive. Here, we present the structure of the diterpene synthase CotB2, in complex with an in crystallo cyclised abrupt reaction product and a substrate-derived diphosphate. We captured additional snapshots of the reaction to gain an overview of CotB2’s catalytic mechanism. To enhance insights into catalysis, structural information is augmented with multiscale molecular dynamic simulations. Our data represent fundamental TPS structure dynamics during catalysis, which ultimately enable rational engineering towards tailored terpene macrocycles that are inaccessible by conventional chemical synthesis.

Original languageEnglish
Article number3971
JournalNature Communications
Volume9
Issue number1
DOIs
StatePublished - 28 Sep 2018

Bibliographical note

Publisher Copyright:
© 2018, The Author(s).

Funding

R. Driller is supported by Elsa-Neumann and Nüsslein-Volhard stipends. D.T. Major acknowledges support from the Israel Science Foundation (Grant #2146/15). T. Brück gratefully acknowledges funding by the Werner Siemens foundation for establishing the field of Synthetic Biotechnology at the Technical University of Munich (TUM). We accessed beamlines of the BESSY II (Berliner Elektronenspeicherring-Gesellschaft für Synchrotronstrahlung II) storage ring (Berlin, Germany) via the Joint Berlin MX-Laboratory sponsored by the Helmholtz Zentrum Berlin für Materialien und Energie, the Freie Universität Berlin, the Humboldt-Universität zu Berlin, the Max-Delbrück Centrum and the Leibniz-Institut für Molekulare Pharmakologie. Parts of this research were carried out at PETRA III at DESY, a member of the Helmholtz Association (HGF). We would like to thank A. Burkhardt for assistance in using beamline P11 and G. Bourenkov for the assistance in using beamline P14. We are grateful to M. Wahl for continuous encouragement and support. We acknowledge support by the German Research Foundation and the Open Access Publication Fund of the Freie Universität Berlin and Technische Universität München.

FundersFunder number
Werner Siemens Foundation
Deutsche Forschungsgemeinschaft
Israel Science Foundation2146/15
Freie Universität Berlin

    Fingerprint

    Dive into the research topics of 'Towards a comprehensive understanding of the structural dynamics of a bacterial diterpene synthase during catalysis'. Together they form a unique fingerprint.

    Cite this