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Protein Pyrophosphorylation by Diphosphoinositol Pentakisphosphate (InsP7)

  • J. Kent WernerJr.
  • Traci Speed
  • Rashna Bhandari
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 645)

Abstract

Diphosphoinositol polyphosphates, also known as inositol pyrophosphates, are a family of water soluble inositol phosphates that possess diphosphate or pyrophosphate moieties. In the presence of divalent cations such as Mg2+, the “high energy” β phosphate can be transferred from the inositol pyrophosphates, InsP 7 and InsP 8, to prephosphorylated serine residues on proteins, to form pyrophosphoserine. This chapter provides detailed methods to identify proteins that are substrates for pyrophosphorylation by InsP 7, conduct phosphorylation assays on purified protein, and detect protein pyrophosphorylation.

Key words

Inositol pyrophosphate Diphosphoinositol pentakisphosphate Pyrophosphoserine Pyrophosphorylation Phosphorylation 

Notes

Acknowledgements

We thank Adolfo Saiardi and Solomon H. Snyder for suggestions and helpful comments. This work is supported by the Department of Biotechnology, Government of India, and The Wellcome Trust, UK.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • J. Kent WernerJr.
    • 1
  • Traci Speed
    • 2
  • Rashna Bhandari
    • 3
  1. 1.The Solomon H. Snyder Department of NeuroscienceJohns Hopkins University School of MedicineBaltimoreUSA
  2. 2.Department of Pharmacology and Molecular SciencesJohns Hopkins University School of MedicineBaltimoreUSA
  3. 3.Laboratory of Cell SignalingCentre for DNA Fingerprinting and Diagnostics (CDFD)NampallyIndia

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