Abstract
With the help of single-cell microflorimetry, 45Ca2+ radiotracer fluxes, and patch-clamp in whole-cell configuration, we examined the effect of the amiloride derivative 3-amino-6-chloro-5-[(4-chloro-benzyl)amino]- N-[[(2,4-dimethylbenzyl)amino]-iminomethyl]-pyrazinecarboxamide (CB-DMB) on the activity of the three isoforms of the Na+/Ca2+ exchanger (NCX) and on several other membrane currents including voltage- and pHsensitive ones. This amiloride analog suppressed the bidirectional activity of all NCX isoforms in a concentration-dependent manner. The IC50 values of CB-DMB were in the nanomolar range for the outward and the inward components of the bidirectional NCX1, NCX2, and NCX3 activity. Deletion mutagenesis showed that CB-DMB inhibited NCX activity mainly at level of the f-loop but not through the interaction with Gly833 located at the level of the α2 repeat. On the other hand, CB-DMB suppressed in the micromolar range the other plasma membrane currents encoded by voltage-dependent Ca2+ channels, tetrodotoxin-sensitive Na+ channels, and pH-sensitive ASIC1a. Collectively, the data of the present study showed that CBDMB, when used in the nanomolar range, is one of the most potent compounds that can block the activity of the three NCX isoforms when they work both in the forward and in the reverse modes of operation without interfering with other ionic channels.
| Original language | English |
|---|---|
| Pages (from-to) | 212-221 |
| Number of pages | 10 |
| Journal | Journal of Pharmacology and Experimental Therapeutics |
| Volume | 331 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Oct 2009 |
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ASJC Scopus subject areas
- Pharmacology
- Molecular Medicine
Cite this
Molecular pharmacology of the amiloride analog 3-amino-6-chloro-5-[(4- chloro-benzyl)amino]-N-[[(2,4-dimethylbenzyl)-amino]iminomethyl] -pyrazinecarboxamide (CB-DMB) as a pan inhibitor of the Na+-Ca 2+ exchanger isoforms NCX1, NCX2, and NCX3 in stably transfected cells. / Secondo, Agnese; Pannaccione, Anna; Molinaro, Pasquale; Ambrosino, Paolo; Lippiello, Pellegrino; Esposito, Alba; Cantile, Maria; Khatri, Priti R.; Melisi, Daniela; Di Renzo, Gianfranco; Annunziato, Lucio.
In: Journal of Pharmacology and Experimental Therapeutics, Vol. 331, No. 1, 10.2009, p. 212-221.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Molecular pharmacology of the amiloride analog 3-amino-6-chloro-5-[(4- chloro-benzyl)amino]-N-[[(2,4-dimethylbenzyl)-amino]iminomethyl] -pyrazinecarboxamide (CB-DMB) as a pan inhibitor of the Na+-Ca 2+ exchanger isoforms NCX1, NCX2, and NCX3 in stably transfected cells
AU - Secondo, Agnese
AU - Pannaccione, Anna
AU - Molinaro, Pasquale
AU - Ambrosino, Paolo
AU - Lippiello, Pellegrino
AU - Esposito, Alba
AU - Cantile, Maria
AU - Khatri, Priti R.
AU - Melisi, Daniela
AU - Di Renzo, Gianfranco
AU - Annunziato, Lucio
PY - 2009/10
Y1 - 2009/10
N2 - With the help of single-cell microflorimetry, 45Ca2+ radiotracer fluxes, and patch-clamp in whole-cell configuration, we examined the effect of the amiloride derivative 3-amino-6-chloro-5-[(4-chloro-benzyl)amino]- N-[[(2,4-dimethylbenzyl)amino]-iminomethyl]-pyrazinecarboxamide (CB-DMB) on the activity of the three isoforms of the Na+/Ca2+ exchanger (NCX) and on several other membrane currents including voltage- and pHsensitive ones. This amiloride analog suppressed the bidirectional activity of all NCX isoforms in a concentration-dependent manner. The IC50 values of CB-DMB were in the nanomolar range for the outward and the inward components of the bidirectional NCX1, NCX2, and NCX3 activity. Deletion mutagenesis showed that CB-DMB inhibited NCX activity mainly at level of the f-loop but not through the interaction with Gly833 located at the level of the α2 repeat. On the other hand, CB-DMB suppressed in the micromolar range the other plasma membrane currents encoded by voltage-dependent Ca2+ channels, tetrodotoxin-sensitive Na+ channels, and pH-sensitive ASIC1a. Collectively, the data of the present study showed that CBDMB, when used in the nanomolar range, is one of the most potent compounds that can block the activity of the three NCX isoforms when they work both in the forward and in the reverse modes of operation without interfering with other ionic channels.
AB - With the help of single-cell microflorimetry, 45Ca2+ radiotracer fluxes, and patch-clamp in whole-cell configuration, we examined the effect of the amiloride derivative 3-amino-6-chloro-5-[(4-chloro-benzyl)amino]- N-[[(2,4-dimethylbenzyl)amino]-iminomethyl]-pyrazinecarboxamide (CB-DMB) on the activity of the three isoforms of the Na+/Ca2+ exchanger (NCX) and on several other membrane currents including voltage- and pHsensitive ones. This amiloride analog suppressed the bidirectional activity of all NCX isoforms in a concentration-dependent manner. The IC50 values of CB-DMB were in the nanomolar range for the outward and the inward components of the bidirectional NCX1, NCX2, and NCX3 activity. Deletion mutagenesis showed that CB-DMB inhibited NCX activity mainly at level of the f-loop but not through the interaction with Gly833 located at the level of the α2 repeat. On the other hand, CB-DMB suppressed in the micromolar range the other plasma membrane currents encoded by voltage-dependent Ca2+ channels, tetrodotoxin-sensitive Na+ channels, and pH-sensitive ASIC1a. Collectively, the data of the present study showed that CBDMB, when used in the nanomolar range, is one of the most potent compounds that can block the activity of the three NCX isoforms when they work both in the forward and in the reverse modes of operation without interfering with other ionic channels.
UR - http://www.scopus.com/inward/record.url?scp=70349322593&partnerID=8YFLogxK
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U2 - 10.1124/jpet.109.152132
DO - 10.1124/jpet.109.152132
M3 - Article
C2 - 19602550
AN - SCOPUS:70349322593
VL - 331
SP - 212
EP - 221
JO - Journal of Pharmacology and Experimental Therapeutics
JF - Journal of Pharmacology and Experimental Therapeutics
SN - 0022-3565
IS - 1
ER -