Identification of amino acid networks governing catalysis in the closed complex of class I terpene synthases

Patrick Schrepfer, Alexander Buettner, Christian Goerner, Michael Hertel, Jeaphianne van Rijn, Frank Wallrapp, Wolfgang Eisenreich, Volker Sieber, Robert Kourist, Thomas Brück

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Class I terpene synthases generate the structural core of bioactive terpenoids. Deciphering structure-function relationships in the reactive closed complex and targeted engineering is hampered by highly dynamic carbocation rearrangements during catalysis. Available crystal structures, however, represent the open, catalytically inactive form or harbor nonproductive substrate analogs. Here, we present a catalytically relevant, closed conformation of taxadiene synthase (TXS), the model class I terpene synthase, which simulates the initial catalytic time point. In silico modeling of subsequent catalytic steps allowed unprecedented insights into the dynamic reaction cascades and promiscuity mechanisms of class I terpene synthases. This generally applicable methodology enables the active-site localization of carbocations and demonstrates the presence of an active-site base motif and its dominating role during catalysis. It additionally allowed in silico-designed targeted protein engineering that unlocked the path to alternate monocyclic and bicyclic synthons representing the basis of a myriad of bioactive terpenoids.

Original languageEnglish
Pages (from-to)E958-67
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number8
DOIs
Publication statusPublished - 23 Feb 2016
Externally publishedYes

Fingerprint

Catalysis
Terpenes
Amino Acids
Computer Simulation
Catalytic Domain
Protein Engineering
terpene synthase

Keywords

  • Alkyl and Aryl Transferases
  • Amino Acid Motifs
  • Catalysis
  • Catalytic Domain
  • Models, Molecular
  • Sequence Analysis, Protein
  • Journal Article
  • Research Support, Non-U.S. Gov't

Fields of Expertise

  • Human- & Biotechnology

Cite this

Identification of amino acid networks governing catalysis in the closed complex of class I terpene synthases. / Schrepfer, Patrick; Buettner, Alexander; Goerner, Christian; Hertel, Michael; van Rijn, Jeaphianne; Wallrapp, Frank; Eisenreich, Wolfgang; Sieber, Volker; Kourist, Robert; Brück, Thomas.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, No. 8, 23.02.2016, p. E958-67.

Research output: Contribution to journalArticleResearchpeer-review

Schrepfer, Patrick ; Buettner, Alexander ; Goerner, Christian ; Hertel, Michael ; van Rijn, Jeaphianne ; Wallrapp, Frank ; Eisenreich, Wolfgang ; Sieber, Volker ; Kourist, Robert ; Brück, Thomas. / Identification of amino acid networks governing catalysis in the closed complex of class I terpene synthases. In: Proceedings of the National Academy of Sciences of the United States of America. 2016 ; Vol. 113, No. 8. pp. E958-67.
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AU - van Rijn, Jeaphianne

AU - Wallrapp, Frank

AU - Eisenreich, Wolfgang

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AB - Class I terpene synthases generate the structural core of bioactive terpenoids. Deciphering structure-function relationships in the reactive closed complex and targeted engineering is hampered by highly dynamic carbocation rearrangements during catalysis. Available crystal structures, however, represent the open, catalytically inactive form or harbor nonproductive substrate analogs. Here, we present a catalytically relevant, closed conformation of taxadiene synthase (TXS), the model class I terpene synthase, which simulates the initial catalytic time point. In silico modeling of subsequent catalytic steps allowed unprecedented insights into the dynamic reaction cascades and promiscuity mechanisms of class I terpene synthases. This generally applicable methodology enables the active-site localization of carbocations and demonstrates the presence of an active-site base motif and its dominating role during catalysis. It additionally allowed in silico-designed targeted protein engineering that unlocked the path to alternate monocyclic and bicyclic synthons representing the basis of a myriad of bioactive terpenoids.

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KW - Sequence Analysis, Protein

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