Maurocalcine interacts with the cardiac ryanodine receptor without inducing channel modification.
We have previously shown that MCa (maurocalcine), a toxin from the venom of the scorpion Maurus palmatus, binds to RyR1 (type 1 ryanodine receptor) and induces strong modifications of its gating behaviour. In the present study, we investigated the ability of MCa to bind to and modify the gating process of cardiac RyR2. By performing pull-down experiments we show that MCa interacts directly with RyR2 with an apparent affinity of 150 nM. By expressing different domains of RyR2 in vitro, we show that MCa binds to two domains of RyR2, which are homologous with those previously identified on RyR1. The effect of MCa binding to RyR2 was then evaluated by three different approaches: (i) [(3)H]ryanodine binding experiments, showing a very weak effect of MCa (up to 1 muM), (ii) Ca(2+) release measurements from cardiac sarcoplasmic reticulum vesicles, showing that MCa up to 1 muM is unable to induce Ca(2+) release, and (iii) single-channel recordings, showing that MCa has no effect on the open probability or on the RyR2 channel conductance level. Long-lasting opening events of RyR2 were observed in the presence of MCa only when the ionic current direction was opposite to the physiological direction, i.e. from the cytoplasmic face of RyR2 to its luminal face. Therefore, despite the conserved MCa binding ability of RyR1 and RyR2, functional studies show that, in contrast with what is observed with RyR1, MCa does not affect the gating properties of RyR2. These results highlight a different role of the MCa-binding domains in the gating process of RyR1 and RyR2.
Altafaj
Xavier
X
iRTSV/CCFP CEA Grenoble INSERM U836 Institut des Neurosciences Grenoble GIN, 17 rue des Martyrs, 38054 Grenoble Cedex 09, France.
France
Julien
J
Almassy
Janos
J
Jona
Istvan
I
Rossi
Daniela
D
Sorrentino
Vincenzo
V
Mabrouk
Kamel
K
De Waard
Michel
M
Ronjat
Michel
M
D000818:Animals; D006321:Heart / Q000187:drug effects*; D015640:Ion Channel Gating / Q000187:drug effects; D009206:Myocardium / Q000378:metabolism*; D011485:Protein Binding; D011817:Rabbits; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D012604:Scorpion Venoms / Q000494:pharmacology*
D019837:Ryanodine Receptor Calcium Release Channel; D012604:Scorpion Venoms; C406406:maurocalcine