BRET-monitoring of the dynamic changes of inositol lipid pools in living cells reveals a PKC-dependent PtdIns4P increase upon EGF and M3 receptor activation
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Tartalom: http://dx.doi.org/10.1016/j.bbalip.2015.12.005
Archívum: MTA Könyvtár
Gyűjtemény: Status = Published



Type = Article
Cím:
BRET-monitoring of the dynamic changes of inositol lipid pools in living cells reveals a PKC-dependent PtdIns4P increase upon EGF and M3 receptor activation
Létrehozó:
Tóth, József T.
Gulyás, Gergő
Tóth, Dániel J.
Balla, András
Hammond, Gerard, R. V.
Hunyady, László
Balla, Tamás
Várnai, Péter
Dátum:
2016
Téma:
QH3011 Biochemistry / biokémia
QH3015 Molecular biology / molekuláris biológia
QH3020 Biophysics / biofizika
Tartalmi leírás:
Deciphering many roles played by inositol lipids in signal transduction and membrane function demands experimental approaches that can detect their dynamic accumulation with subcellular accuracy and exquisite sensitivity. The former criterion is met by imaging of fluorescence biosensors in living cells, whereas the latter is facilitated by biochemical measurements from populations. Here, we introduce BRET-based biosensors able to detect rapid changes in inositol lipids in cell populations with both high sensitivity and subcellular resolution in a single, convenient assay. We demonstrate robust and sensitive measurements of PtdIns4P, PtdIns(4,5)P2 and PtdIns(3,4,5)P3 dynamics, as well as changes in cytoplasmic Ins(1,4,5)P3 levels. Measurements were made during either experimental activation of lipid degradation, or PI 3-kinase and phospholipase C mediated signal transduction. Our results reveal a previously unappreciated synthesis of PtdIns4P that accompanies moderate activation of phospholipase C signaling downstream of both EGF and muscarinic M3 receptor activation. This signaling-induced PtdIns4P synthesis relies on protein kinase C, and implicates a feedback mechanism in the control of inositol lipid metabolism during signal transduction. © 2015 Elsevier B.V. All rights reserved.
Típus:
Article
PeerReviewed
Formátum:
text
Azonosító:
Tóth, József T. and Gulyás, Gergő and Tóth, Dániel J. and Balla, András and Hammond, Gerard, R. V. and Hunyady, László and Balla, Tamás and Várnai, Péter (2016) BRET-monitoring of the dynamic changes of inositol lipid pools in living cells reveals a PKC-dependent PtdIns4P increase upon EGF and M3 receptor activation. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS, 1861 (3). pp. 177-187. ISSN 1388-1981
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