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Kinetics of Extracellular ATP in Mastoparan 7-activated Human Erythrocytes

Kinetics of Extracellular ATP in Mastoparan 7-activated Human Erythrocytes

María Florencia Leal Denis, J Jeremías Incicco, María Victoria Espelt, Sandra V Verstraeten, Omar P Pignataro, Eduardo R Lazarowski, Pablo J Schwarzbaum

Biochim Biophys Acta. 2013 Oct;1830(10):4692-707.

PMID: 23742824

Abstract:

Background:
The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intra- and extracellular processes governing the time-dependent accumulation of extracellular ATP (i.e., ATPe kinetics).
Methods:
Human erythrocytes were treated with MST7 in the presence or absence of two blockers of pannexin 1. ATPe concentration was monitored by luciferin-luciferase based real-time luminometry.
Results:
Exposure of human erythrocytes to MST7 led to an acute increase in [ATPe], followed by a slower increase phase. ATPe kinetics reflected a strong activation of ATP efflux and a low rate of ATPe hydrolysis by ectoATPase activity. Enhancement of [ATPe] by MST7 required adhesion of erythrocytes to poly-D-lysin-coated coverslips, and correlated with a 31% increase of cAMP and 10% cell swelling. However, when MST7 was dissolved in a hyperosmotic medium to block cell swelling, ATPe accumulation was inhibited by 49%. Erythrocytes pre-exposure to 10μM of either carbenoxolone or probenecid, two blockers of pannexin 1, exhibited a partial reduction of ATP efflux. Erythrocytes from pannexin 1 knockout mice exhibited similar ATPe kinetics as those of wild type mice erythrocytes exposed to pannexin 1 blockers.
Conclusions:
MST7 induced release of ATP required either cell adhesion or strong activation of cAMP synthesis. Part of this release required cell swelling. Kinetic analysis and a data driven model suggested that ATP efflux is mediated by two ATP conduits displaying different kinetics, with one conduit being fully blocked by pannexin 1 blockers.
General significance:
Kinetic analysis of extracellular ATP accumulation from human erythrocytes and potential effects on microcirculation.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP145854597	Mastoparan-7		145854-59-7	Price

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