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- Title
Inositol Pyrophosphates and Their Unique Metabolic Complexity: Analysis by Gel Electrophoresis.
- Authors
Losito, Oriana; Szijgyarto, Zsolt; Resnick, Adam Cain; Saiardi, Adolfo
- 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.
- Subjects
INOSITOL phosphates; PYROPHOSPHATES; POLYACRYLAMIDE gel electrophoresis; TOLUIDINE blue; HIGH performance liquid chromatography; CELL metabolism; CELLULAR control mechanisms
- Publication
PLoS ONE, 2009, Vol 4, Issue 5, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0005580