Atomic resolution structures of discrete stages on the reaction coordinate of the [Fe4S4] enzyme IspG (GcpE)

Handle URI:
http://hdl.handle.net/10754/550086
Title:
Atomic resolution structures of discrete stages on the reaction coordinate of the [Fe4S4] enzyme IspG (GcpE)
Authors:
Quitterer, Felix; Frank, Annika; Wang, Ke; Rao, Guodong; O'Dowd, Bing; Li, Jikun; Guerra, Francisco; Abdel-Azeim, Safwat; Bacher, Adelbert; Eppinger, Jörg; Oldfield, Eric; Groll, Michael
Abstract:
IspG is the penultimate enzyme in non-mevalonate biosynthesis of the universal terpene building blocks isopentenyl diphosphate and dimethylallyl diphosphate. Its mechanism of action has been the subject of numerous studies but remained unresolved due to difficulties in identifying distinct reaction intermediates. Using a moderate reducing agent as well as an epoxide substrate analogue, we were now able to trap and crystallographically characterize various stages in the IspG catalyzed conversion of 2-C-methyl-D-erythritol-2,4-cyclo-diphosphate (MEcPP) to (E)-1-hydroxy-2-methylbut-2-enyl-4-diphosphate (HMBPP). In addition, the enzyme’s structure was determined in complex with several inhibitors. These results, combined with recent electron paramagnetic resonance data, allowed us to deduce a detailed and complete IspG catalytic mechanism which describes all stages from initial ring opening to formation of HMBPP via discrete radical and carbanion intermediates. The data presented in this article provide a guide for the design of selective drugs against many pro- and eukaryotic pathogens to which the non-mevalonate pathway is essential for survival and virulence.
KAUST Department:
Physical Sciences and Engineering (PSE) Division; KAUST Catalysis Center (KCC)
Citation:
Quitterer, Felix, Annika Frank, Ke Wang, Guodong Rao, Bing O'Dowd, Jikun Li, Francisco Guerra et al. "Atomic resolution structures of discrete stages on the reaction coordinate of the [Fe4S4] enzyme IspG (GcpE)." Journal of Molecular Biology (2015).
Publisher:
Elsevier
Journal:
Journal of Molecular Biology
Issue Date:
11-Apr-2015
DOI:
10.1016/j.jmb.2015.04.002
PubMed ID:
25868383
PubMed Central ID:
PMC4433817
Type:
Article
ISSN:
00222836
Additional Links:
http://linkinghub.elsevier.com/retrieve/pii/S0022283615002259
Appears in Collections:
Articles; Physical Sciences and Engineering (PSE) Division; KAUST Catalysis Center (KCC)

Full metadata record

DC FieldValue Language
dc.contributor.authorQuitterer, Felixen
dc.contributor.authorFrank, Annikaen
dc.contributor.authorWang, Keen
dc.contributor.authorRao, Guodongen
dc.contributor.authorO'Dowd, Bingen
dc.contributor.authorLi, Jikunen
dc.contributor.authorGuerra, Franciscoen
dc.contributor.authorAbdel-Azeim, Safwaten
dc.contributor.authorBacher, Adelberten
dc.contributor.authorEppinger, Jörgen
dc.contributor.authorOldfield, Ericen
dc.contributor.authorGroll, Michaelen
dc.date.accessioned2015-04-14T12:38:16Zen
dc.date.available2015-04-14T12:38:16Zen
dc.date.issued2015-04-11en
dc.identifier.citationQuitterer, Felix, Annika Frank, Ke Wang, Guodong Rao, Bing O'Dowd, Jikun Li, Francisco Guerra et al. "Atomic resolution structures of discrete stages on the reaction coordinate of the [Fe4S4] enzyme IspG (GcpE)." Journal of Molecular Biology (2015).en
dc.identifier.issn00222836en
dc.identifier.pmid25868383en
dc.identifier.doi10.1016/j.jmb.2015.04.002en
dc.identifier.urihttp://hdl.handle.net/10754/550086en
dc.description.abstractIspG is the penultimate enzyme in non-mevalonate biosynthesis of the universal terpene building blocks isopentenyl diphosphate and dimethylallyl diphosphate. Its mechanism of action has been the subject of numerous studies but remained unresolved due to difficulties in identifying distinct reaction intermediates. Using a moderate reducing agent as well as an epoxide substrate analogue, we were now able to trap and crystallographically characterize various stages in the IspG catalyzed conversion of 2-C-methyl-D-erythritol-2,4-cyclo-diphosphate (MEcPP) to (E)-1-hydroxy-2-methylbut-2-enyl-4-diphosphate (HMBPP). In addition, the enzyme’s structure was determined in complex with several inhibitors. These results, combined with recent electron paramagnetic resonance data, allowed us to deduce a detailed and complete IspG catalytic mechanism which describes all stages from initial ring opening to formation of HMBPP via discrete radical and carbanion intermediates. The data presented in this article provide a guide for the design of selective drugs against many pro- and eukaryotic pathogens to which the non-mevalonate pathway is essential for survival and virulence.en
dc.publisherElsevieren
dc.relation.urlhttp://linkinghub.elsevier.com/retrieve/pii/S0022283615002259en
dc.rightsNOTICE: this is the author’s version of a work that was accepted for publication in Journal of Molecular Biology. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Journal of Molecular Biology, 11 April 2015. DOI: 10.1016/j.jmb.2015.04.002en
dc.subjectterpene biosynthesisen
dc.subjectmethylerythritol-phosphate pathwayen
dc.subjectiron-sulfur cluster catalysisen
dc.subjectreaction mechanismsen
dc.titleAtomic resolution structures of discrete stages on the reaction coordinate of the [Fe4S4] enzyme IspG (GcpE)en
dc.typeArticleen
dc.contributor.departmentPhysical Sciences and Engineering (PSE) Divisionen
dc.contributor.departmentKAUST Catalysis Center (KCC)en
dc.identifier.journalJournal of Molecular Biologyen
dc.identifier.pmcidPMC4433817en
dc.eprint.versionPost-printen
dc.contributor.institutionCenter for Integrated Protein Science, Department Chemie, Lehrstuhl für Biochemie, Technische Universität München,Garching D-85747, Germanyen
dc.contributor.institutionDepartment of Chemistry, University of Illinois, Urbana, IL 61801, United Statesen
kaust.authorAbdel-Azeim, Safwaten
kaust.authorEppinger, Jorgen

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