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PLoS One. 2009;4(5):e5580. doi: 10.1371/journal.pone.0005580. Epub 2009 May 18.

Inositol pyrophosphates and their unique metabolic complexity: analysis by gel electrophoresis.

Author information

1
Department of Cell and Developmental Biology, Medical Research Council Cell Biology Unit and Laboratory for Molecular Cell Biology, University College London, London, United States of America.

Abstract

BACKGROUND:

Inositol pyrophosphates are a recently characterized cell signalling molecules responsible for the pyrophosphorylation of protein substrates. Though likely involved in a wide range of cellular functions, the study of inositol pyrophosphates has suffered from a lack of readily available methods for their analysis.

PRINCIPAL FINDING:

We describe a novel, sensitive and rapid polyacrylamide gel electrophoresis (PAGE)-based method for the analysis of inositol pyrophosphates. Using 4',6-diamidino-2-phenylindole (DAPI) and Toluidine Blue we demonstrate the unequivocal detection of various inositol pyrophosphate species.

CONCLUSION:

The use of the PAGE-based method reveals the likely underestimation of inositol pyrophosphates and their signalling contribution in cells when measured via traditional HPLC-based techniques. PAGE-based analyses also reveals the existence of a number of additional, previously uncharacterised pyrophosphorylated inositol reaction products, defining a more complex metabolism associated with the catalytically flexible kinase class responsible for the production of these highly energetic cell signalling molecules.

PMID:
19440344
PMCID:
PMC2680042
DOI:
10.1371/journal.pone.0005580
[Indexed for MEDLINE]
Free PMC Article

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