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

<h4>Background</h4>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...

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Main Authors: Oriana Losito, Zsolt Szijgyarto, Adam Cain Resnick, Adolfo Saiardi
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2009-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0005580
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author Oriana Losito
Zsolt Szijgyarto
Adam Cain Resnick
Adolfo Saiardi
author_facet Oriana Losito
Zsolt Szijgyarto
Adam Cain Resnick
Adolfo Saiardi
author_sort Oriana Losito
collection DOAJ
description <h4>Background</h4>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.<h4>Principal finding</h4>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.<h4>Conclusion</h4>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.
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spelling doaj-art-a2ab1b545456456ba879d04c4cdcec1b2025-08-20T02:00:52ZengPublic Library of Science (PLoS)PLoS ONE1932-62032009-01-0145e558010.1371/journal.pone.0005580Inositol pyrophosphates and their unique metabolic complexity: analysis by gel electrophoresis.Oriana LositoZsolt SzijgyartoAdam Cain ResnickAdolfo Saiardi<h4>Background</h4>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.<h4>Principal finding</h4>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.<h4>Conclusion</h4>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.https://doi.org/10.1371/journal.pone.0005580
spellingShingle Oriana Losito
Zsolt Szijgyarto
Adam Cain Resnick
Adolfo Saiardi
Inositol pyrophosphates and their unique metabolic complexity: analysis by gel electrophoresis.
PLoS ONE
title Inositol pyrophosphates and their unique metabolic complexity: analysis by gel electrophoresis.
title_full Inositol pyrophosphates and their unique metabolic complexity: analysis by gel electrophoresis.
title_fullStr Inositol pyrophosphates and their unique metabolic complexity: analysis by gel electrophoresis.
title_full_unstemmed Inositol pyrophosphates and their unique metabolic complexity: analysis by gel electrophoresis.
title_short Inositol pyrophosphates and their unique metabolic complexity: analysis by gel electrophoresis.
title_sort inositol pyrophosphates and their unique metabolic complexity analysis by gel electrophoresis
url https://doi.org/10.1371/journal.pone.0005580
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