Skip to main content
Springer Nature Link
Log in

Farnesyl Diphosphate Synthase Assay

  • Protocol
  • First Online:
Plant Isoprenoids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1153))

Abstract

Farnesyl diphosphate synthase (FPS) catalyzes the sequential head-to-tail condensation of isopentenyl diphosphate (IPP, C5) with dimethylallyl diphosphate (DMAPP, C5) and geranyl diphosphate (GPP, C10) to produce farnesyl diphosphate (FPP, C15). This short-chain prenyl diphosphate constitutes a key branch-point of the isoprenoid biosynthetic pathway from which a variety of bioactive isoprenoids that are vital for normal plant growth and survival are produced. Here we describe a protocol to obtain highly purified preparations of recombinant FPS and a radiochemical assay method for measuring FPS activity in purified enzyme preparations and plant tissue extracts.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Kellogg BA, Poulter CD (1997) Chain elongation in the isoprenoid biosynthetic pathway. Curr Opin Chem Biol 1:570–578

    Article  CAS  PubMed  Google Scholar 

  2. Poulter CD (2006) Farnesyl diphosphate synthase. A paradigm for understanding structure and function relationships in E-polyprenyl diphosphate synthases. Phytochem Rev 5:17–26

    Article  CAS  Google Scholar 

  3. Dhar MK, Koul A, Kaul S (2013) Farnesyl pyrophosphate synthase: a key enzyme in isoprenoid biosynthetic pathway and potential molecular target for drug development. New Biotechnol 30:114–123

    Article  CAS  Google Scholar 

  4. Koyama T, Gotoh Y, Nishino T (2000) Intersubunit location of the active site of farnesyl diphosphate synthase: reconstruction of active enzymes by hybrid-type heteromeric dimers of site-directed mutants. Biochemistry 39:463–469

    Article  CAS  PubMed  Google Scholar 

  5. Ambo T, Noike M, Kurokawa H, Koyama T (2008) Cloning and functional analysis of novel short-chain cis-prenyltransferases. Biochem Biophys Res Commun 375:536–540

    Article  CAS  PubMed  Google Scholar 

  6. Sallaud C, Rontein D, Onillon S, Jabès F, Duffé P, Giacalone C, Thoraval S, Escoffier C, Herbette G, Leonhardt N, Causse M, Tissier A (2009) A novel pathway for sesquiterpene biosynthesis from Z,Z-farnesyl pyrophosphate in the wild tomato Solanum habrochaites. Plant Cell 21:301–317

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Tholl D, Sungbeon L (2011) Terpene specialized metabolism in Arabidopsis thaliana. Arabidopsis Book 9:e0143. doi:10.1043/tab.0143

    Article  PubMed Central  PubMed  Google Scholar 

  8. Keim V, Manzano D, Fernández FJ, Closa M, Andrade P, Caudepón D, Bortolotti C, Vega MC, Arró M, Ferrer A (2012) Characterization of Arabidopsis FPS isozymes and FPS gene expression analysis provide insight into the biosynthesis of isoprenoid precursors in seeds. PLoS ONE 7:e49109. doi:10.1371/journal.pone.0049109.t001

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. Cunillera N, Boronat A, Ferrer A (1997) The Arabidopsis thaliana FPS1 gene generates a novel mRNA that encodes a mitochondrial farnesyl-diphosphate synthase isoform. J Biol Chem 272:15381–15388

    Article  CAS  PubMed  Google Scholar 

  10. Thabet I, Guirimand G, Courdavault V, Papon N, Godet S, Dutilleul C, Bouzid S, Giglioli-Guivarc'h N, Clastre M, Simkin AJ (2011) The subcellular localization of periwinkle farnesyl diphosphate synthase provides insight into the role of peroxisome in isoprenoid biosynthesis. J Plant Physiol 168:2110–2116

    Article  CAS  PubMed  Google Scholar 

  11. Sanmiya K, Ueno O, Matsuoka M, Yamamoto N (1999) Localization of farnesyl diphosphate synthase in chloroplasts. Plant Cell Physiol 40:348–354

    Article  CAS  PubMed  Google Scholar 

  12. Cunillera N, Arró M, Delourme D, Karst F, Boronat A, Ferrer A (1996) Arabidopsis thaliana contains two differentially expressed farnesyl-diphosphate synthase genes. J Biol Chem 271:7774–7780

    Article  CAS  PubMed  Google Scholar 

  13. Brinkmann U, Mattes RE, Buckel P (1989) High-level expression of recombinant genes in Escherichia coli is dependent on the availability of the dnaY gene product. Gene 85:109–114

    Article  CAS  PubMed  Google Scholar 

  14. Mekkriengkrai D, Sando T, Hirooka K, Sakdapipanich J, Tanaka Y, Fukusaki E, Kobayashi A (2004) Cloning and characterization of farnesyl diphosphate synthase from the rubber-producing mushroom Lactarius chrysorrheus. Biosci Biotechnol Biochem 68:2360–2368

    Article  CAS  PubMed  Google Scholar 

  15. Sen SE, Trobaugh C, Béliveau C, Richard T, Cusson M (2007) Cloning, expression and characterization of a dipteran farnesyl diphosphate synthase. Insect Biochem Mol Biol 37:1198–1206

    Article  CAS  PubMed  Google Scholar 

  16. Ogura K, Nishino T, Shinka T, Seto S (1985) Prenyltransferases of pumpkin fruit. Methods Enzymol 110:167–171

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the Spanish Ministerio de Ciencia e Innovación (BIO2009-06984, cofinanced by the European Regional Development Funds), the Spanish Consolider-Ingenio 2010 Program (CSD2007-00036, Center for Research in Agricultural Genomics), and the Generalitat de Catalunya (2009SGR0026).

Author information

Authors and Affiliations

  1. Center for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Campus UAB Bellaterra, Barcelona, Spain

    Montserrat Arró, David Manzano & Albert Ferrer

  2. Faculty of Pharmacy, Department of Biochemistry and Molecular Biology, University of Barcelona, DIAGONAL, 643 08028, Barcelona, Spain

    Montserrat Arró

  3. Faculty of Pharmacy, Department of Biochemistry and Molecular Biology, University of Barcelona, DIAGONAL, 643 08028, Barcelona, Spain

    David Manzano & Albert Ferrer

Authors
  1. Montserrat Arró
    View author publications

    Search author on:PubMed Google Scholar

  2. David Manzano
    View author publications

    Search author on:PubMed Google Scholar

  3. Albert Ferrer
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Albert Ferrer .

Editor information

Editors and Affiliations

  1. Centre for Research in Agricultural Genomic (CRAG), CSIC-IRTA-UAB-UB, Barcelona, Spain

    Manuel Rodríguez-Concepción

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Arró, M., Manzano, D., Ferrer, A. (2014). Farnesyl Diphosphate Synthase Assay. In: Rodríguez-Concepción, M. (eds) Plant Isoprenoids. Methods in Molecular Biology, vol 1153. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0606-2_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-0606-2_4

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0605-5

  • Online ISBN: 978-1-4939-0606-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics