File Download
 
Links for fulltext
(May Require Subscription)
 
Supplementary

Article: Epidermal growth factor receptor tyrosine kinase regulates the human inward rectifier potassium K IR2.3 channel, stably expressed in HEK 293 cells
  • Basic View
  • Metadata View
  • XML View
TitleEpidermal growth factor receptor tyrosine kinase regulates the human inward rectifier potassium K IR2.3 channel, stably expressed in HEK 293 cells
 
AuthorsZhang, DY1
Zhang, YH1
Sun, HY1
Lau, CP1
Li, GR1 1
 
KeywordsEGFR tyrosine kinase
inward rectifier K + channels
protein tyrosine kinase
protein tyrosine phosphatase
tyrosine phosphorylation
 
Issue Date2011
 
PublisherJohn Wiley & Sons Ltd. The Journal's web site is located at http://www.wiley.com/bw/journal.asp?ref=0007-1188&site=1
 
CitationBritish Journal Of Pharmacology, 2011, v. 164 n. 5, p. 1469-1478 [How to Cite?]
DOI: http://dx.doi.org/10.1111/j.1476-5381.2011.01424.x
 
AbstractBackground and Purpose The detailed molecular modulation of inward rectifier potassium channels (including the K IR2.3 channel) is not fully understood. The present study was designed to determine whether human K IR2.3 (K IR2.3) channels were regulated by protein tyrosine kinases (PTKs). Experimental Approach Whole-cell patch voltage-clamp, immunoprecipitation, Western blot analysis and site-directed mutagenesis were employed to determine the potential PTK phosphorylation of Kir2.3 current in HEK 293 cells stably expressing Kir2.3 gene. Key Results The broad-spectrum PTK inhibitor genistein (10 ÂμM) and the selective epidermal growth factor (EGF) kinase inhibitor AG556 (10 ÂμM) reversibly decreased K IR2.3 current and the effect was reversed by the protein tyrosine phosphatase inhibitor, orthovanadate (1 mM). Although EGF (100 ng·mL -1) and orthovanadate enhanced K IR2.3 current, this effect was antagonized by AG556. However, the Src-family tyrosine kinase inhibitor PP2 (10 ÂμM) did not inhibit K IR2.3 current. Tyrosine phosphorylation of K IR2.3 channels was decreased by genistein or AG556, and was increased by EGF or orthovanadate. The decrease of tyrosine phosphorylation of K IR2.3 channels by genistein or AG556 was reversed by orthovanadate or EGF. Interestingly, the response of K IR2.3 channels to EGF or AG556 was lost in the K IR2.3 Y234A mutant channel. Conclusion and Implications These results demonstrate that the EGF receptor tyrosine kinase up-regulates the K IR2.3 channel via phosphorylation of the Y234 residue of the WT protein. This effect may be involved in the endogenous regulation of cellular electrical activity. © 2011 The British Pharmacological Society.
 
ISSN0007-1188
2012 Impact Factor: 5.067
2012 SCImago Journal Rankings: 1.488
 
DOIhttp://dx.doi.org/10.1111/j.1476-5381.2011.01424.x
 
PubMed Central IDPMC3221101
 
ISI Accession Number IDWOS:000296472300009
Funding AgencyGrant Number
Research Grant Council of Hong KongHKU 760306 M
Sun Chieh Yeh Heart Foundation of Hong Kong
University of Hong Kong
Funding Information:

This work was supported in part by a General Research Fund (HKU 760306 M) from Research Grant Council of Hong Kong and a grant from Sun Chieh Yeh Heart Foundation of Hong Kong. The authors greatly appreciate the generous offer of pcDNA3.1/hK<INF>IR</INF>2.3 plasmid for the present study by Dr Carol A Vandenberg, University of California Santa Barbara. De-Yong Zhang and Yan-Hui Zhang are supported by post-graduate studentships from the University of Hong Kong.

 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorZhang, DY
 
dc.contributor.authorZhang, YH
 
dc.contributor.authorSun, HY
 
dc.contributor.authorLau, CP
 
dc.contributor.authorLi, GR
 
dc.date.accessioned2011-10-28T02:45:11Z
 
dc.date.available2011-10-28T02:45:11Z
 
dc.date.issued2011
 
dc.description.abstractBackground and Purpose The detailed molecular modulation of inward rectifier potassium channels (including the K IR2.3 channel) is not fully understood. The present study was designed to determine whether human K IR2.3 (K IR2.3) channels were regulated by protein tyrosine kinases (PTKs). Experimental Approach Whole-cell patch voltage-clamp, immunoprecipitation, Western blot analysis and site-directed mutagenesis were employed to determine the potential PTK phosphorylation of Kir2.3 current in HEK 293 cells stably expressing Kir2.3 gene. Key Results The broad-spectrum PTK inhibitor genistein (10 ÂμM) and the selective epidermal growth factor (EGF) kinase inhibitor AG556 (10 ÂμM) reversibly decreased K IR2.3 current and the effect was reversed by the protein tyrosine phosphatase inhibitor, orthovanadate (1 mM). Although EGF (100 ng·mL -1) and orthovanadate enhanced K IR2.3 current, this effect was antagonized by AG556. However, the Src-family tyrosine kinase inhibitor PP2 (10 ÂμM) did not inhibit K IR2.3 current. Tyrosine phosphorylation of K IR2.3 channels was decreased by genistein or AG556, and was increased by EGF or orthovanadate. The decrease of tyrosine phosphorylation of K IR2.3 channels by genistein or AG556 was reversed by orthovanadate or EGF. Interestingly, the response of K IR2.3 channels to EGF or AG556 was lost in the K IR2.3 Y234A mutant channel. Conclusion and Implications These results demonstrate that the EGF receptor tyrosine kinase up-regulates the K IR2.3 channel via phosphorylation of the Y234 residue of the WT protein. This effect may be involved in the endogenous regulation of cellular electrical activity. © 2011 The British Pharmacological Society.
 
dc.description.naturelink_to_OA_fulltext
 
dc.identifier.citationBritish Journal Of Pharmacology, 2011, v. 164 n. 5, p. 1469-1478 [How to Cite?]
DOI: http://dx.doi.org/10.1111/j.1476-5381.2011.01424.x
 
dc.identifier.doihttp://dx.doi.org/10.1111/j.1476-5381.2011.01424.x
 
dc.identifier.epage1478
 
dc.identifier.hkuros197656
 
dc.identifier.isiWOS:000296472300009
Funding AgencyGrant Number
Research Grant Council of Hong KongHKU 760306 M
Sun Chieh Yeh Heart Foundation of Hong Kong
University of Hong Kong
Funding Information:

This work was supported in part by a General Research Fund (HKU 760306 M) from Research Grant Council of Hong Kong and a grant from Sun Chieh Yeh Heart Foundation of Hong Kong. The authors greatly appreciate the generous offer of pcDNA3.1/hK<INF>IR</INF>2.3 plasmid for the present study by Dr Carol A Vandenberg, University of California Santa Barbara. De-Yong Zhang and Yan-Hui Zhang are supported by post-graduate studentships from the University of Hong Kong.

 
dc.identifier.issn0007-1188
2012 Impact Factor: 5.067
2012 SCImago Journal Rankings: 1.488
 
dc.identifier.issue5
 
dc.identifier.pmcidPMC3221101
 
dc.identifier.pmid21486282
 
dc.identifier.scopuseid_2-s2.0-80054043745
 
dc.identifier.spage1469
 
dc.identifier.urihttp://hdl.handle.net/10722/142403
 
dc.identifier.volume164
 
dc.languageeng
 
dc.publisherJohn Wiley & Sons Ltd. The Journal's web site is located at http://www.wiley.com/bw/journal.asp?ref=0007-1188&site=1
 
dc.publisher.placeUnited Kingdom
 
dc.relation.ispartofBritish Journal of Pharmacology
 
dc.relation.referencesReferences in Scopus
 
dc.rightsBritish Journal of Pharmacology. Copyright © John Wiley & Sons Ltd.
 
dc.subject.meshBlotting, Western
 
dc.subject.meshElectrophysiology
 
dc.subject.meshGenistein - pharmacology
 
dc.subject.meshPotassium Channels, Inwardly Rectifying - genetics - metabolism
 
dc.subject.meshReceptor, Epidermal Growth Factor - antagonists and inhibitors - physiology
 
dc.subjectEGFR tyrosine kinase
 
dc.subjectinward rectifier K + channels
 
dc.subjectprotein tyrosine kinase
 
dc.subjectprotein tyrosine phosphatase
 
dc.subjecttyrosine phosphorylation
 
dc.titleEpidermal growth factor receptor tyrosine kinase regulates the human inward rectifier potassium K IR2.3 channel, stably expressed in HEK 293 cells
 
dc.typeArticle
 
<?xml encoding="utf-8" version="1.0"?>
<item><contributor.author>Zhang, DY</contributor.author>
<contributor.author>Zhang, YH</contributor.author>
<contributor.author>Sun, HY</contributor.author>
<contributor.author>Lau, CP</contributor.author>
<contributor.author>Li, GR</contributor.author>
<date.accessioned>2011-10-28T02:45:11Z</date.accessioned>
<date.available>2011-10-28T02:45:11Z</date.available>
<date.issued>2011</date.issued>
<identifier.citation>British Journal Of Pharmacology, 2011, v. 164 n. 5, p. 1469-1478</identifier.citation>
<identifier.issn>0007-1188</identifier.issn>
<identifier.uri>http://hdl.handle.net/10722/142403</identifier.uri>
<description.abstract>Background and Purpose The detailed molecular modulation of inward rectifier potassium channels (including the K IR2.3 channel) is not fully understood. The present study was designed to determine whether human K IR2.3 (K IR2.3) channels were regulated by protein tyrosine kinases (PTKs). Experimental Approach Whole-cell patch voltage-clamp, immunoprecipitation, Western blot analysis and site-directed mutagenesis were employed to determine the potential PTK phosphorylation of Kir2.3 current in HEK 293 cells stably expressing Kir2.3 gene. Key Results The broad-spectrum PTK inhibitor genistein (10 &#194;&#956;M) and the selective epidermal growth factor (EGF) kinase inhibitor AG556 (10 &#194;&#956;M) reversibly decreased K IR2.3 current and the effect was reversed by the protein tyrosine phosphatase inhibitor, orthovanadate (1 mM). Although EGF (100 ng&#183;mL -1) and orthovanadate enhanced K IR2.3 current, this effect was antagonized by AG556. However, the Src-family tyrosine kinase inhibitor PP2 (10 &#194;&#956;M) did not inhibit K IR2.3 current. Tyrosine phosphorylation of K IR2.3 channels was decreased by genistein or AG556, and was increased by EGF or orthovanadate. The decrease of tyrosine phosphorylation of K IR2.3 channels by genistein or AG556 was reversed by orthovanadate or EGF. Interestingly, the response of K IR2.3 channels to EGF or AG556 was lost in the K IR2.3 Y234A mutant channel. Conclusion and Implications These results demonstrate that the EGF receptor tyrosine kinase up-regulates the K IR2.3 channel via phosphorylation of the Y234 residue of the WT protein. This effect may be involved in the endogenous regulation of cellular electrical activity. &#169; 2011 The British Pharmacological Society.</description.abstract>
<language>eng</language>
<publisher>John Wiley &amp; Sons Ltd. The Journal&apos;s web site is located at http://www.wiley.com/bw/journal.asp?ref=0007-1188&amp;site=1</publisher>
<relation.ispartof>British Journal of Pharmacology</relation.ispartof>
<rights>British Journal of Pharmacology. Copyright &#169; John Wiley &amp; Sons Ltd.</rights>
<subject>EGFR tyrosine kinase</subject>
<subject>inward rectifier K + channels</subject>
<subject>protein tyrosine kinase</subject>
<subject>protein tyrosine phosphatase</subject>
<subject>tyrosine phosphorylation</subject>
<subject.mesh>Blotting, Western</subject.mesh>
<subject.mesh>Electrophysiology</subject.mesh>
<subject.mesh>Genistein - pharmacology</subject.mesh>
<subject.mesh>Potassium Channels, Inwardly Rectifying - genetics - metabolism</subject.mesh>
<subject.mesh>Receptor, Epidermal Growth Factor - antagonists and inhibitors - physiology</subject.mesh>
<title>Epidermal growth factor receptor tyrosine kinase regulates the human inward rectifier potassium K IR2.3 channel, stably expressed in HEK 293 cells</title>
<type>Article</type>
<description.nature>link_to_OA_fulltext</description.nature>
<identifier.doi>10.1111/j.1476-5381.2011.01424.x</identifier.doi>
<identifier.pmid>21486282</identifier.pmid>
<identifier.pmcid>PMC3221101</identifier.pmcid>
<identifier.scopus>eid_2-s2.0-80054043745</identifier.scopus>
<identifier.hkuros>197656</identifier.hkuros>
<relation.references>http://www.scopus.com/mlt/select.url?eid=2-s2.0-80054043745&amp;selection=ref&amp;src=s&amp;origin=recordpage</relation.references>
<identifier.volume>164</identifier.volume>
<identifier.issue>5</identifier.issue>
<identifier.spage>1469</identifier.spage>
<identifier.epage>1478</identifier.epage>
<identifier.isi>WOS:000296472300009</identifier.isi>
<publisher.place>United Kingdom</publisher.place>
<bitstream.url>http://hub.hku.hk/bitstream/10722/142403/1/re01.htm</bitstream.url>
</item>
Author Affiliations
  1. The University of Hong Kong Li Ka Shing Faculty of Medicine