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

Losito, Oriana

Resnick, Adam Cain

Saiardi, Adolfo

Szijgyarto, Zsolt

English

PubMed

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.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.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

Losito Oriana

Szijgyarto Zsolt

Resnick Adam Cain

Saiardi Adolfo

Public Library of Science (PLOS)

Inositol Pyrophosphates and Their Unique Metabolic Complexity: Analysis by Gel Electrophoresis

Losito, O

Szijgyarto, Z

Resnick, AC

Saiardi, A

UCL Discovery

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

BackgroundInositol 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 findingWe 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.ConclusionThe 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

Oriana Losito

Zsolt Szijgyarto

Adam Cain Resnick

Adolfo Saiardi

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PLoS ONE

https://figshare.com/articles/Inositol_Pyrophosphates_and_Their_Unique_Metabolic_Complexity_Analysis_by_Gel_Electrophoresis/147575

Inositol Pyrophosphates and Their Unique Metabolic Complexity: Analysis by Gel Electrophoresis

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UCL Discovery

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