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


RESULTS
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.


CONCLUSIONS
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.
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.
Oriana
O
Losito
Zsolt
Z
Szijgyarto
Adam Cain
AC
Resnick
Adolfo
A
Saiardi
D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D007295:Inositol Phosphates / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
D007295:Inositol Phosphates