A novel mass assay to quantify the bioactive lipid PtdIns3P in various biological samples.

Gaëtan Chicanne 1 Sonia Severin 1 Cécile Boscheron 2 Anne-Dominique Terrisse 1 Marie-Pierre Gratacap 1 Frédérique Gaits-Iacovoni 1 Hélène Tronchère 1 Bernard Payrastre 1, 3, *
* Auteur correspondant
1 I2MC - Institut des Maladies Métaboliques et Cardiovasculaires
2 INSERM U836, équipe 1, Physiopathologie du cytosquelette
GPC-GIN - Groupe Physiopathologie du Cytosquelette, GIN - Grenoble Institut des Neurosciences
3 Laboratoire d'Hématologie [Purpan]
Abstract : PtdIns3P is recognized as an important player in the control of the endocytotic pathway and in autophagy. Recent data also suggest that PtdIns3P contributes to molecular mechanisms taking place at the plasma membrane and at the midbody during cytokinesis. This lipid is present in low amounts in mammalian cells and remains difficult to quantify either by traditional techniques based on radiolabelling followed by HPLC to separate the different phosphatidylinositol monophosphates, or by high-sensitive liquid chromatography coupled to MS, which is still under development. In the present study, we describe a mass assay to quantify this lipid from various biological samples using the recombinant PtdIns3P 5-kinase, PIKfyve. Using this assay, we show an increase in the mass level of PtdIns3P in mouse and human platelets following stimulation, loss of this lipid in Vps34-deficient yeasts and its relative enrichment in early endosomes isolated from BHK cells.
keyword : class III phosphoinositide 3-kinase mass assay PIKfyve PtdIns3P
Type de document :
Article dans une revue
Biochemical Journal, Portland Press, 2012, 447 (1), pp.17-23. 〈10.1042/BJ20120945〉
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Gaëtan Chicanne, Sonia Severin, Cécile Boscheron, Anne-Dominique Terrisse, Marie-Pierre Gratacap, et al.. A novel mass assay to quantify the bioactive lipid PtdIns3P in various biological samples.. Biochemical Journal, Portland Press, 2012, 447 (1), pp.17-23. 〈10.1042/BJ20120945〉. 〈inserm-00739444〉

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