Article: Modulation of human cardiac transient outward potassium current by EGFR tyrosine kinase and Src-family kinases
| Title | Modulation of human cardiac transient outward potassium current by EGFR tyrosine kinase and Src-family kinases |
|---|---|
| Authors | Zhang, YH1 Wu, W1 Sun, HY1 Deng, XL2 Cheng, LC1 Li, X1 Tse, HF1 Lau, CP1 Li, GR1 |
| Keywords | EGFR kinase hKv4.3 Human atrial myocytes Protein tyrosine kinases Protein tyrosine phosphatase Src-family kinases Transient outward potassium current |
| Issue Date | 2012 |
| Publisher | Oxford University Press. The Journal's web site is located at http://cardiovascres.oxfordjournals.org |
| Citation | Cardiovascular Research, 2012, v. 93 n. 3, p. 424-433 [How to Cite?] DOI: http://dx.doi.org/10.1093/cvr/cvr347 |
| Abstract | Aims: The human cardiac transient outward K + current Ito (encoded by Kv4.3 or KCND3) plays an important role in phase 1 rapid repolarization of cardiac action potentials in the heart. However, modulation of I to by intracellular signal transduction is not fully understood. The present study was therefore designed to determine whether/how human atrial I to and hKv4.3 channels stably expressed in HEK 293 cells are regulated by protein tyrosine kinases (PTKs). Methods and results: Whole-cell patch voltage-clamp, immunoprecipitation, western blotting, and site-directed mutagenesis approaches were employed in the present study. We found that human atrial I to was inhibited by the broad-spectrum PTK inhibitor genistein, the selective epidermal growth factor receptor (EGFR) kinase inhibitor AG556, and the Src-family kinases inhibitor PP2. The inhibitory effect was countered by the protein tyrosine phosphatase inhibitor orthovanadate. In HEK 293 cells stably expressing human KCND3, genistein, AG556, and PP2 significantly reduced the hKv4.3 current, and the reduction was antagonized by orthovanadate. Interestingly, orthovanadate also reversed the reduced tyrosine phosphorylation level of hKv4.3 channels by genistein, AG556, or PP2. Mutagenesis revealed that the hKv4.3 mutant Y136F lost the inhibitory response to AG556, while Y108F lost response to PP2. The double-mutant Y108FY136F hKv4.3 channels showed no response to either AG556 or PP2. Conclusion: Our results demonstrate that human atrial Ito and cloned hKv4.3 channels are modulated by EGFR kinase via phosphorylation of the Y136 residue and by Src-family kinases via phosphorylation of the Y108 residue; tyrosine phosphorylation of the channel may be involved in regulating cardiac electrophysiology. © The Author 2011. |
| ISSN | 0008-6363 2011 Impact Factor: 6.064 2011 SCImago Journal Rankings: 0.649 |
| DOI | http://dx.doi.org/10.1093/cvr/cvr347 |
| References | References in Scopus |
| dc.contributor.author | Zhang, YH | ||||||
|---|---|---|---|---|---|---|---|
| dc.contributor.author | Wu, W | ||||||
| dc.contributor.author | Sun, HY | ||||||
| dc.contributor.author | Deng, XL | ||||||
| dc.contributor.author | Cheng, LC | ||||||
| dc.contributor.author | Li, X | ||||||
| dc.contributor.author | Tse, HF | ||||||
| dc.contributor.author | Lau, CP | ||||||
| dc.contributor.author | Li, GR | ||||||
| dc.date.accessioned | 2012-02-03T06:13:38Z | ||||||
| dc.date.available | 2012-02-03T06:13:38Z | ||||||
| dc.date.issued | 2012 | ||||||
| dc.description.abstract | Aims: The human cardiac transient outward K + current Ito (encoded by Kv4.3 or KCND3) plays an important role in phase 1 rapid repolarization of cardiac action potentials in the heart. However, modulation of I to by intracellular signal transduction is not fully understood. The present study was therefore designed to determine whether/how human atrial I to and hKv4.3 channels stably expressed in HEK 293 cells are regulated by protein tyrosine kinases (PTKs). Methods and results: Whole-cell patch voltage-clamp, immunoprecipitation, western blotting, and site-directed mutagenesis approaches were employed in the present study. We found that human atrial I to was inhibited by the broad-spectrum PTK inhibitor genistein, the selective epidermal growth factor receptor (EGFR) kinase inhibitor AG556, and the Src-family kinases inhibitor PP2. The inhibitory effect was countered by the protein tyrosine phosphatase inhibitor orthovanadate. In HEK 293 cells stably expressing human KCND3, genistein, AG556, and PP2 significantly reduced the hKv4.3 current, and the reduction was antagonized by orthovanadate. Interestingly, orthovanadate also reversed the reduced tyrosine phosphorylation level of hKv4.3 channels by genistein, AG556, or PP2. Mutagenesis revealed that the hKv4.3 mutant Y136F lost the inhibitory response to AG556, while Y108F lost response to PP2. The double-mutant Y108FY136F hKv4.3 channels showed no response to either AG556 or PP2. Conclusion: Our results demonstrate that human atrial Ito and cloned hKv4.3 channels are modulated by EGFR kinase via phosphorylation of the Y136 residue and by Src-family kinases via phosphorylation of the Y108 residue; tyrosine phosphorylation of the channel may be involved in regulating cardiac electrophysiology. © The Author 2011. | ||||||
| dc.description.nature | postprint | ||||||
| dc.identifier.citation | Cardiovascular Research, 2012, v. 93 n. 3, p. 424-433 [How to Cite?] DOI: http://dx.doi.org/10.1093/cvr/cvr347 | ||||||
| dc.identifier.doi | http://dx.doi.org/10.1093/cvr/cvr347 | ||||||
| dc.identifier.epage | 433 | ||||||
| dc.identifier.hkuros | 198242 | ||||||
| dc.identifier.isi | WOS:000300789300009
Funding Information: The work was supported in part by a grant from Sun Chieh Yeh Heart Foundation of Hong Kong. Y.H.Z. and W.W. were supported by a post-graduate scholarship from the University of Hong Kong. | ||||||
| dc.identifier.issn | 0008-6363 2011 Impact Factor: 6.064 2011 SCImago Journal Rankings: 0.649 | ||||||
| dc.identifier.issue | 3 | ||||||
| dc.identifier.pmid | 22198508 | ||||||
| dc.identifier.scopus | eid_2-s2.0-84863276532 | ||||||
| dc.identifier.spage | 424 | ||||||
| dc.identifier.uri | http://hdl.handle.net/10722/144554 | ||||||
| dc.identifier.volume | 93 | ||||||
| dc.language | eng | ||||||
| dc.publisher | Oxford University Press. The Journal's web site is located at http://cardiovascres.oxfordjournals.org | ||||||
| dc.publisher.place | United Kingdom | ||||||
| dc.relation.ispartof | Cardiovascular Research | ||||||
| dc.relation.references | References in Scopus | ||||||
| dc.rights | This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Cardiovascular Research following peer review. The definitive publisher-authenticated version Cardiovascular Research, 2012, v. 93 n. 3, p. 424-433 is available online at: http://cardiovascres.oxfordjournals.org/content/93/3/424 | ||||||
| dc.rights | Creative Commons: Attribution 3.0 Hong Kong License | ||||||
| dc.subject | EGFR kinase | ||||||
| dc.subject | hKv4.3 | ||||||
| dc.subject | Human atrial myocytes | ||||||
| dc.subject | Protein tyrosine kinases | ||||||
| dc.subject | Protein tyrosine phosphatase | ||||||
| dc.subject | Src-family kinases | ||||||
| dc.subject | Transient outward potassium current | ||||||
| dc.title | Modulation of human cardiac transient outward potassium current by EGFR tyrosine kinase and Src-family kinases | ||||||
| dc.type | Article |
Author Affiliations
- The University of Hong Kong Li Ka Shing Faculty of Medicine
- Xi'an Jiaotong University, School of Medicine