Involvment of protein tyrosine phosphorylation and chloride channel in transient receptor potential protein mediated storeoperated calcium influx

Qin Ying Qiu, Xiao R. Yang, Hua He, Jing Liang Li, Xue Rong Wang, Yong Yuan Guan

Research output: Contribution to journalArticle

Abstract

AIM: To investigate whether protein tyrosine phosphorylation and Cl- channels be involved in transient receptor potential(TRP) protein mediated store-operated calcium influx(SOC) in HEK293 cells. METHODS: Human TRP1 (hTRP1) and human TRP3(hTRP3) cDNA were transfected to HEK293 cells respectively using lipofectAMINE reagent, the effects of genistein, furosemide and 4, 4-diisothiocyanostilbene-2, 2-disulphonic acid(DIDS) on thapsigargin (TG)-induced Ca2+ influx were determined by Fura-2/AM spectrophoto-fluorometry. RESULTS: Compared with untransfected cells, the significant enhancement in TG-induced Ca2+ influx in hTRP1-transfected HEK293 cells was found, but not in hTRP3-transfected ones. Genistein 5 - 30 μmol · L-1, furosemide 1 - 8 μmol · L-1 and DIDS 0.5 - 1.0 μmol · L-1 inhibited TG-induced Ca2+ influx in hTRP1-transfected cells. CONCLUSION: hTRP1 Protein may be one of components of TG-induced SOC in transfected HEK293 cells. Tyrosine phosphorylation and Cl- channels may be involved in TG-induced Ca2+ influx and tyrosine kinase may regulate hTRP1 protein directly.

Original languageEnglish (US)
Pages (from-to)172-178
Number of pages7
JournalChinese Journal of Pharmacology and Toxicology
Volume17
Issue number3
StatePublished - Jun 2003
Externally publishedYes

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Chloride Channels
Phosphorylation
Thapsigargin
HEK293 Cells
Tyrosine
Calcium
Genistein
Furosemide
Proteins
Fluorometry
Acids
Protein-Tyrosine Kinases
Complementary DNA

Keywords

  • 4,4-diisothiocyanostilbene-2,2-disulphonic acid
  • Calcium channels
  • Chloride channels
  • Furosemide
  • Genistein
  • Protein-tyrosine kinase
  • Store-operated calcium influx
  • Transient receptor potential protein

ASJC Scopus subject areas

  • Pharmacology
  • Toxicology

Cite this

Involvment of protein tyrosine phosphorylation and chloride channel in transient receptor potential protein mediated storeoperated calcium influx. / Qiu, Qin Ying; Yang, Xiao R.; He, Hua; Li, Jing Liang; Wang, Xue Rong; Guan, Yong Yuan.

In: Chinese Journal of Pharmacology and Toxicology, Vol. 17, No. 3, 06.2003, p. 172-178.

Research output: Contribution to journalArticle

Qiu, Qin Ying ; Yang, Xiao R. ; He, Hua ; Li, Jing Liang ; Wang, Xue Rong ; Guan, Yong Yuan. / Involvment of protein tyrosine phosphorylation and chloride channel in transient receptor potential protein mediated storeoperated calcium influx. In: Chinese Journal of Pharmacology and Toxicology. 2003 ; Vol. 17, No. 3. pp. 172-178.
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AU - Wang, Xue Rong

AU - Guan, Yong Yuan

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N2 - AIM: To investigate whether protein tyrosine phosphorylation and Cl- channels be involved in transient receptor potential(TRP) protein mediated store-operated calcium influx(SOC) in HEK293 cells. METHODS: Human TRP1 (hTRP1) and human TRP3(hTRP3) cDNA were transfected to HEK293 cells respectively using lipofectAMINE reagent, the effects of genistein, furosemide and 4, 4-diisothiocyanostilbene-2, 2-disulphonic acid(DIDS) on thapsigargin (TG)-induced Ca2+ influx were determined by Fura-2/AM spectrophoto-fluorometry. RESULTS: Compared with untransfected cells, the significant enhancement in TG-induced Ca2+ influx in hTRP1-transfected HEK293 cells was found, but not in hTRP3-transfected ones. Genistein 5 - 30 μmol · L-1, furosemide 1 - 8 μmol · L-1 and DIDS 0.5 - 1.0 μmol · L-1 inhibited TG-induced Ca2+ influx in hTRP1-transfected cells. CONCLUSION: hTRP1 Protein may be one of components of TG-induced SOC in transfected HEK293 cells. Tyrosine phosphorylation and Cl- channels may be involved in TG-induced Ca2+ influx and tyrosine kinase may regulate hTRP1 protein directly.

AB - AIM: To investigate whether protein tyrosine phosphorylation and Cl- channels be involved in transient receptor potential(TRP) protein mediated store-operated calcium influx(SOC) in HEK293 cells. METHODS: Human TRP1 (hTRP1) and human TRP3(hTRP3) cDNA were transfected to HEK293 cells respectively using lipofectAMINE reagent, the effects of genistein, furosemide and 4, 4-diisothiocyanostilbene-2, 2-disulphonic acid(DIDS) on thapsigargin (TG)-induced Ca2+ influx were determined by Fura-2/AM spectrophoto-fluorometry. RESULTS: Compared with untransfected cells, the significant enhancement in TG-induced Ca2+ influx in hTRP1-transfected HEK293 cells was found, but not in hTRP3-transfected ones. Genistein 5 - 30 μmol · L-1, furosemide 1 - 8 μmol · L-1 and DIDS 0.5 - 1.0 μmol · L-1 inhibited TG-induced Ca2+ influx in hTRP1-transfected cells. CONCLUSION: hTRP1 Protein may be one of components of TG-induced SOC in transfected HEK293 cells. Tyrosine phosphorylation and Cl- channels may be involved in TG-induced Ca2+ influx and tyrosine kinase may regulate hTRP1 protein directly.

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