The canonical transient receptor potential 6 channel as a putative phosphatidylinositol 3,4,5-trisphosphate-sensitive calcium entry system

Ping Hui Tseng, Ho Pi Lin, Hongzhen Hu, Chunbo Wang, Michael Xi Zhu, Ching Shih Chen

Research output: Contribution to journalArticle

74 Scopus citations

Abstract

We previously reported that phosphatidylinositol 3,4,5-trisphosphate (PIP3), a lipid product of phosphoinositide 3-kinase (PI3K), induced Ca2+ influx via a noncapacitative pathway in platelets, Jurkat T cells, and RBL-2H3 mast cells. The identity of this Ca2+ influx system, however, remains unclear. Here, we investigate a potential link between PIP3-sensitive Ca2+ entry and the canonical transient receptor potential (TRPC) channels by developing stable human embryonic kidney (HEK) 293 cell lines expressing TRPC1, TRPC3, TRPC5, and TRPC6. Two lines of evidence support TRPC6 as a putative target by which PIP3 induces Ca2+ influx. First, Fura-2 fluorometric Ca2+ analysis shows the ability of PIP3 to selectively stimulate [Ca 2+]i increase in TRPC6-expressing cells. Second, pull-down analysis indicates specific interactions between biotin-PIP3 and TRPC6 protein. Our data indicate that PIP3 activates store-independent Ca2+ entry in TRPC6 cells via a nonselective cation channel. Although the activating effect of PIP3 on TRPC6 is reminiscent to that of 1-oleoyl-2-acetyl-sn-glycerol, this activation is not attributable to the diacylglycerol substructure of PIP3 since other phosphoinositides failed to trigger Ca2+ responses. The PIP 3-activated Ca2+ entry is inhibited by known TRPC6 inhibitors such as Gd3+ and SKF96365 and is independent of IP 3 production. Furthermore, we demonstrated that TRPC6 overexpression or antisense downregulation significantly alters the amplitude of PIP 3- and anti-CD3-activated Ca2+ responses in Jurkat T cells. Consequently, the link between TRPC6 and PIP3-mediated Ca 2+ entry provides a framework to account for an intimate relationship between PI3K and PLCγ in initiating Ca2+ response to agonist stimulation in T lymphocytes.

Original languageEnglish
Pages (from-to)11701-11708
Number of pages8
JournalBiochemistry
Volume43
Issue number37
DOIs
StatePublished - Sep 21 2004
Externally publishedYes

Fingerprint Dive into the research topics of 'The canonical transient receptor potential 6 channel as a putative phosphatidylinositol 3,4,5-trisphosphate-sensitive calcium entry system'. Together they form a unique fingerprint.

  • Cite this