Abstract
Na+/Ca2+ exchanger (NCX) is a plasma membrane transporter that moves Ca2+ in or out of the cell, depending on membrane potential and transmembrane ion gradients. NCX is the main pathway for Ca2+ extrusion from excitable cells. NCX inhibitors can ameliorate cardiac ischemia-reperfusion injury and promote high-frequency fatigue of skeletal muscle, purportedly by inhibiting the Ca2+ inward mode of NCX. Here we tested two known NCX inhibitors, 2-(2-(4-(4-nitrobenzyloxy)phenyl) ethyl)-isothiourea methanesulfonate (KB-R7943) and the structurally related 2-[[4-[(4-Nitrophenyl)methoxy]phenyl]methyl]-4-thiazoli dinecarboxylic acid ethyl ester (SN-6), for their influence on electrically or caffeine-evoked Ca2+ transients in adult dissociated flexor digitorum brevis (FDB) skeletal muscle fibers and human embryonic kidney (HEK) 293 cells that have stable expression of type 1 ryanodine receptor (RyR1). KB-R7943 (≤10 μM) reversibly attenuates electrically evoked Ca2+ transients in FDB and caffeine-induced Ca2+ release in HEK 293, whereas the structurally related NCX inhibitor SN-6 does not, suggesting that KB-R7943 directly inhibits RyR1. In support of this interpretation, KB-R7943 inhibits high-affinity binding of [3H]ryanodine to RyR1 (IC50 = 5.1 ± 0.9 μM) and the cardiac isoform RyR2 (IC50 = 13.4 ± 1.8 μM). KB-R7943 interfered with the gating of reconstituted RyR1 and RyR2 channels, reducing open probability (Po), shortening mean open time, and prolonging mean closed time. KB-R7943 was more effective at blocking RyR1 with cytoplasmic conditions favoring high Po compared with those favoring low Po. SN-6 has negligible activity toward altering [ 3H]ryanodine binding of RyR1 and RyR2. Our results identify that KB-R7943 is a reversible, activity-dependent blocker of the two most broadly expressed RyR channel isoforms and contributes to its pharmacological and therapeutic activities.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 560-568 |
| Number of pages | 9 |
| Journal | Molecular Pharmacology |
| Volume | 76 |
| Issue number | 3 |
| DOIs | |
| State | Published - Sep 2009 |
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ASJC Scopus subject areas
- Pharmacology
- Molecular Medicine
Cite this
The Na+/Ca2+ exchange inhibitor 2-(2-(4-(4- nitrobenzyloxy)phenyl)ethyl)isothiourea methanesulfonate (KB-R7943) also blocks ryanodine receptors type 1 (RyR1) and type 2 (RyR2) channels. / Barrientos, Genaro; Bose, Diptiman D.; Feng, Wei; Padilla, Isela; Pessah, Isaac N.
In: Molecular Pharmacology, Vol. 76, No. 3, 09.2009, p. 560-568.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - The Na+/Ca2+ exchange inhibitor 2-(2-(4-(4- nitrobenzyloxy)phenyl)ethyl)isothiourea methanesulfonate (KB-R7943) also blocks ryanodine receptors type 1 (RyR1) and type 2 (RyR2) channels
AU - Barrientos, Genaro
AU - Bose, Diptiman D.
AU - Feng, Wei
AU - Padilla, Isela
AU - Pessah, Isaac N
PY - 2009/9
Y1 - 2009/9
N2 - Na+/Ca2+ exchanger (NCX) is a plasma membrane transporter that moves Ca2+ in or out of the cell, depending on membrane potential and transmembrane ion gradients. NCX is the main pathway for Ca2+ extrusion from excitable cells. NCX inhibitors can ameliorate cardiac ischemia-reperfusion injury and promote high-frequency fatigue of skeletal muscle, purportedly by inhibiting the Ca2+ inward mode of NCX. Here we tested two known NCX inhibitors, 2-(2-(4-(4-nitrobenzyloxy)phenyl) ethyl)-isothiourea methanesulfonate (KB-R7943) and the structurally related 2-[[4-[(4-Nitrophenyl)methoxy]phenyl]methyl]-4-thiazoli dinecarboxylic acid ethyl ester (SN-6), for their influence on electrically or caffeine-evoked Ca2+ transients in adult dissociated flexor digitorum brevis (FDB) skeletal muscle fibers and human embryonic kidney (HEK) 293 cells that have stable expression of type 1 ryanodine receptor (RyR1). KB-R7943 (≤10 μM) reversibly attenuates electrically evoked Ca2+ transients in FDB and caffeine-induced Ca2+ release in HEK 293, whereas the structurally related NCX inhibitor SN-6 does not, suggesting that KB-R7943 directly inhibits RyR1. In support of this interpretation, KB-R7943 inhibits high-affinity binding of [3H]ryanodine to RyR1 (IC50 = 5.1 ± 0.9 μM) and the cardiac isoform RyR2 (IC50 = 13.4 ± 1.8 μM). KB-R7943 interfered with the gating of reconstituted RyR1 and RyR2 channels, reducing open probability (Po), shortening mean open time, and prolonging mean closed time. KB-R7943 was more effective at blocking RyR1 with cytoplasmic conditions favoring high Po compared with those favoring low Po. SN-6 has negligible activity toward altering [ 3H]ryanodine binding of RyR1 and RyR2. Our results identify that KB-R7943 is a reversible, activity-dependent blocker of the two most broadly expressed RyR channel isoforms and contributes to its pharmacological and therapeutic activities.
AB - Na+/Ca2+ exchanger (NCX) is a plasma membrane transporter that moves Ca2+ in or out of the cell, depending on membrane potential and transmembrane ion gradients. NCX is the main pathway for Ca2+ extrusion from excitable cells. NCX inhibitors can ameliorate cardiac ischemia-reperfusion injury and promote high-frequency fatigue of skeletal muscle, purportedly by inhibiting the Ca2+ inward mode of NCX. Here we tested two known NCX inhibitors, 2-(2-(4-(4-nitrobenzyloxy)phenyl) ethyl)-isothiourea methanesulfonate (KB-R7943) and the structurally related 2-[[4-[(4-Nitrophenyl)methoxy]phenyl]methyl]-4-thiazoli dinecarboxylic acid ethyl ester (SN-6), for their influence on electrically or caffeine-evoked Ca2+ transients in adult dissociated flexor digitorum brevis (FDB) skeletal muscle fibers and human embryonic kidney (HEK) 293 cells that have stable expression of type 1 ryanodine receptor (RyR1). KB-R7943 (≤10 μM) reversibly attenuates electrically evoked Ca2+ transients in FDB and caffeine-induced Ca2+ release in HEK 293, whereas the structurally related NCX inhibitor SN-6 does not, suggesting that KB-R7943 directly inhibits RyR1. In support of this interpretation, KB-R7943 inhibits high-affinity binding of [3H]ryanodine to RyR1 (IC50 = 5.1 ± 0.9 μM) and the cardiac isoform RyR2 (IC50 = 13.4 ± 1.8 μM). KB-R7943 interfered with the gating of reconstituted RyR1 and RyR2 channels, reducing open probability (Po), shortening mean open time, and prolonging mean closed time. KB-R7943 was more effective at blocking RyR1 with cytoplasmic conditions favoring high Po compared with those favoring low Po. SN-6 has negligible activity toward altering [ 3H]ryanodine binding of RyR1 and RyR2. Our results identify that KB-R7943 is a reversible, activity-dependent blocker of the two most broadly expressed RyR channel isoforms and contributes to its pharmacological and therapeutic activities.
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U2 - 10.1124/mol.109.057265
DO - 10.1124/mol.109.057265
M3 - Article
C2 - 19509218
AN - SCOPUS:69549119075
VL - 76
SP - 560
EP - 568
JO - Molecular Pharmacology
JF - Molecular Pharmacology
SN - 0026-895X
IS - 3
ER -