Abstract
Stimulation of α1-adrenoreceptors (α1-AR) acutely alters ion channel behavior via several signaling pathways [calcium and protein kinase C (PKC)]. Little is known about sustained α1-adrenergic/PKC signaling and channel regulation as may occur during cardiovascular disease states. Here we describe the effects of prolonged α1A-AR and PKC activity on human ether-a-go-go-related gene (HERG) K+ channels (Kv11.1) expressed in a heterologous expression system. Stimulation of α1A-AR with phenylephrine or direct activation of PKC with phorbol ester increased HERG channel protein abundance and K+ current density in a time- and dose-dependent manner. Channel augmentation reached a steady-state plateau within 24 h with a 2- to 6-fold induction. Phorbol ester and moderate α1A-AR stimulation enhanced HERG abundance in a PKC-dependent fashion but with stronger α1A-adrenergic stimulation; protein kinase A (PKA)-dependent activity also contributed. Comparable channel induction of other cardiac K+ channels was not seen in this system. Comparison of wild-type HERG and channels with either mutated PKC phosphorylation sites (HERGΔPKC) or mutated PKA phosphorylation sites (HERGΔPKA) suggested that the mechanisms of augmentation of HERG by the two kinases were partially overlapping. The PKC-dependent effect was largely due to enhanced synthetic rates. Stimulation of α1-AR in cultured rat neonatal cardiac myocytes also enhanced the abundance of ERG channels. These findings show that α1A-AR stimulation is capable of influencing the balance of HERG channel synthesis and degradation via multiple signaling pathways, a process that may have relevance in cardiac diseases and treatment.
Footnotes
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This work was supported by the National Institutes of Health Division of Blood Diseases and Resources [Grant HL077326]; the Singapore Biomedical Research Council; the University of Heidelberg (Frontiers Program); and the Adumed Foundation.
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Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.
doi:10.1124/mol.109.062216.
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ABBREVIATIONS:
- HERG
- human ether-a-go-go-related gene
- ERG
- ether-a-go-go
- HEK
- human embryonic kidney
- PKA
- protein kinase A
- PKC
- protein kinase C
- PMA
- phorbol 12-myristate 13-acetate
- CPT
- chlorophenylthiol
- α1-AR
- α1-adrenoreceptors
- HERGΔPKC
- human ether-a-go-go-related gene with 17 of 18 possible protein kinase C target sites mutated to alanine
- HERGΔPKA
- human ether-a-go-go-related gene with all four possible protein kinas A target sites mutated to alanine
- PE
- phenylephrine
- PBS
- phosphate-buffered saline
- PAGE
- polyacrylamide gel electrophoresis
- TBS
- Tris-buffered saline
- PCR
- polymerase chain reaction
- PIP2
- phosphatidylinositol bisphosphate
- ER
- endoplasmic reticulum
- Hsp90
- 90-kDa heat shock protein
- 5-MU
- 5-methylurapidil
- Gö6976
- 12-(2-cyanoethyl)-6,7,12,13-tetrahydro-13-methyl-5-oxo-5H-indolo(2,3-a)pyrrolo(3,4-c)-carbazole
- Ro 32-0432
- 3-[(8S)-8-[(dimethylamino)methyl]-6,7,8,9-tetrahydropyrido[1,2-a]indol-10-yl]-4-(1-methyl-1H-indol-3-yl)-1H-pyrrole-2,5-dione hydrochloride
- H89
- N-[2-(4-bromocinnamylamino)ethyl]-5-isoquinoline
- LY294002
- 2-(4-morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one hydrochloride
- Bis-I
- bisindolylmaleimide I
- BAPTA
- 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid
- Hsc70
- 70-kDa heat shock protein cognate.
- Received November 6, 2009.
- Accepted May 11, 2010.
- Copyright © 2010 The American Society for Pharmacology and Experimental Therapeutics
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