Regulation of TRP channel TRPM2 by the tyrosine phosphatase PTPL1

Wenyi Zhang, Qin Tong, Kathleen Conrad, Jocelyn Wozney, Joseph Y. Cheung, Barbara Miller

Research output: Contribution to journalArticle

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Abstract

TRPM2, a member of the transient receptor potential (TRP) superfamily, is a Ca2+-permeable channel, which mediates susceptibility to cell death following activation by oxidative stress, TNFα, or β-amyloid peptide. We determined that TRPM2 is rapidly tyrosine phosphorylated after stimulation with H2O2 or TNFα. Inhibition of tyrosine phosphorylation with the tyrosine kinase inhibitors genistein or PP2 significantly reduced the increase in [Ca2+]i observed after H2O2 or TNFα treatment in TRPM2-expressing cells, suggesting that phosphorylation is important in TRPM2 activation. Utilizing a TransSignal PDZ domain array blot to identify proteins which interact with TRPM2, we identified PTPL1 as a potential binding protein. PTPL1 is a widely expressed tyrosine phosphatase, which has a role in cell survival and tumorigenesis. Immunoprecipitation and glutathione-S-transferase pull-down assays confirmed that TRPM2 and PTPL1 interact. To examine the ability of PTPL1 to modulate phosphorylation or activation of TRPM2, PTPL1 was coexpressed with TRPM2 in human embryonic kidney-293T cells. This resulted in significantly reduced TRPM2 tyrosine phosphorylation, and inhibited the rise in [Ca 2+]i and the loss of cell viability, which follow H 2O2 or TNFα treatment. Consistent with these findings, reduction in endogenous PTPL1 expression with small interfering RNA resulted in increased TRPM2 tyrosine phosphorylation, a significantly greater rise in [Ca2+]i following H2O2 treatment, and enhanced susceptibility to H2O2-induced cell death. Endogenous TRPM2 and PTPL1 was associated in U937-ecoR cells, confirming the physiological relevance of this interaction. These data demonstrate that tyrosine phosphorylation of TRPM2 is important in its activation and function and that inhibition of TRPM2 tyrosine phosphorylation reduces Ca2+ influx and protects cell viability. They also suggest that modulation of TRPM2 tyrosine phosphorylation is a mechanism through which PTPL1 may mediate resistance to cell death.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume292
Issue number5
DOIs
StatePublished - May 1 2007

Fingerprint

Transient Receptor Potential Channels
Phosphorylation
Phosphoric Monoester Hydrolases
Tyrosine
Cell death
Chemical activation
Cell Survival
Cell Death
Cells
PDZ Domains
U937 Cells
Oxidative stress
Genistein
HEK293 Cells
Glutathione Transferase
Immunoprecipitation
Amyloid
Protein-Tyrosine Kinases
Small Interfering RNA
Assays

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Zhang, Wenyi ; Tong, Qin ; Conrad, Kathleen ; Wozney, Jocelyn ; Cheung, Joseph Y. ; Miller, Barbara. / Regulation of TRP channel TRPM2 by the tyrosine phosphatase PTPL1. In: American Journal of Physiology - Cell Physiology. 2007 ; Vol. 292, No. 5.
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Regulation of TRP channel TRPM2 by the tyrosine phosphatase PTPL1. / Zhang, Wenyi; Tong, Qin; Conrad, Kathleen; Wozney, Jocelyn; Cheung, Joseph Y.; Miller, Barbara.

In: American Journal of Physiology - Cell Physiology, Vol. 292, No. 5, 01.05.2007.

Research output: Contribution to journalArticle

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AU - Zhang, Wenyi

AU - Tong, Qin

AU - Conrad, Kathleen

AU - Wozney, Jocelyn

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