2 C-methyl-d-erythritol 4-phosphate enhances and sustains cyclodiphosphate synthase IspF activity

J. Kipchirchir Bitok, Caren L Meyers

Research output: Contribution to journalArticle

Abstract

There is significant progress toward understanding catalysis throughout the essential MEP pathway to isoprenoids in human pathogens; however, little is known about pathway regulation. The present study begins by testing the hypothesis that isoprenoid biosynthesis is regulated via feedback inhibition of the fifth enzyme cyclodiphosphate synthase IspF by downstream isoprenoid diphosphates. Here, we demonstrate recombinant E. coli IspF is not inhibited by downstream metabolites isopentenyl diphosphate (IDP), dimethylallyl diphosphate (DMADP), geranyl diphosphate (GDP), and farnesyl diphosphate (FDP) under standard assay conditions. However, 2C-methyl-d-erythritol 4-phosphate (MEP), the product of reductoisomerase IspC and first committed MEP pathway intermediate, activates and sustains this enhanced IspF activity, and the IspF-MEP complex is inhibited by FDP. We further show that the methylerythritol scaffold itself, which is unique to this pathway, drives the activation and stabilization of active IspF. Our results suggest a novel feed-forward regulatory mechanism for 2C-methyl-d-erythritol 2,4-cyclodiphosphate (MEcDP) production and support an isoprenoid biosynthesis regulatory mechanism via feedback inhibition of the IspF-MEP complex by FDP. The results have important implications for development of inhibitors against the IspF-MEP complex, which may be the physiologically relevant form of the enzyme.

Original languageEnglish (US)
Pages (from-to)1702-1710
Number of pages9
JournalACS Chemical Biology
Volume7
Issue number10
DOIs
StatePublished - Oct 19 2012

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Terpenes
Biosynthesis
Erythritol
Feedback
Diphosphates
Pathogens
Enzymes
Metabolites
Catalysis
Scaffolds
Escherichia coli
erythritol 4-phosphate
2-C-methylerythritol 4-phosphate
Assays
Stabilization
Chemical activation
Testing
farnesyl pyrophosphate

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Medicine(all)

Cite this

2 C-methyl-d-erythritol 4-phosphate enhances and sustains cyclodiphosphate synthase IspF activity. / Kipchirchir Bitok, J.; Meyers, Caren L.

In: ACS Chemical Biology, Vol. 7, No. 10, 19.10.2012, p. 1702-1710.

Research output: Contribution to journalArticle

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