Inositol pyrophosphates and their unique metabolic complexity: analysis by gel electrophoresis.
Inositol pyrophosphates and their unique metabolic complexity: analysis by gel electrophoresis.
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DOI：
10.1371/journal.pone.0005580
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Saiardi A
中科院分区：
文献类型：
--
作者：
Losito O;Szijgyarto Z;Resnick AC;Saiardi A
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.
