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Content Provider | Journal of Biological Chemistry (JBC) |
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Author | Choi, Seok Kim, Hyun-Ju Ko, Yoo-Seung Jeong, Seong-Woo Kim, Yang In Simonds, William F. Oh, Jae-Wook Nah, Seung-Yeol |
Abstract | Recently we demonstrated that ginsenosides, the active ingredients of Panax ginseng, enhanced Ca2+-activated Cl− current in theXenopus oocyte through a signal transduction mechanism involving the activation of pertussis toxin-insensitive G protein and phospholipase C (PLC). However, it has not yet been determined precisely which G protein subunit(s) and which PLC isoform(s) participate in the ginsenoside signaling. To provide answers to these questions, we investigated the changes in ginsenoside effect on the Cl− current after intraoocyte injections of the cRNAs coding various G protein subunits, a regulator of G protein signaling (RGS2), and Gβγ-binding proteins. In addition, we examined which of mammalian PLCβ1–3 antibodies injected into the oocyte inhibited the action of ginsenosides on the Cl− current. Injection of Gαq or Gα11 cRNA increased the basal Cl− current recorded 48 h after, and it further prevented ginsenosides from enhancing the Cl− current, whereas Gαi2 and GαoA cRNA injection had no significant effect. The changes following Gαq cRNA injection were prevented when Gβ1γ2 and Gαq subunits were co-expressed by simultaneous injection of the cRNAs coding these subunits. Injection of cRNA coding GαqQ209L, a constitutively active mutant that does not bind to Gβγ, produced effects similar to those of GαqcRNA injection. The effects of GαqQ209L cRNA injection, however, were not prevented by co-injection of Gβ1γ2 cRNA. Injection of the cRNA coding RGS2, which interacts most selectively with Gαq/11 among various identified RGS isoforms and stimulates the hydrolysis of GTP to GDP in active GTP-bound Gα subunit, resulted in a severe attenuation of ginsenoside effect on the Cl− current. Finally, antibodies against PLCβ3, but not -β1 and -β2, markedly attenuated the ginsenoside effect examined at 3-h postinjection. These results suggest that Gαq/11 coupled to mammalian PLC β3-like enzyme mediates ginsenoside effect on Ca2+-activated Cl− current in theXenopus oocyte. |
Related Links | http://www.jbc.org/content/276/52/48797.abstract |
Ending Page | 48802 |
Starting Page | 48797 |
Page Count | 6 |
File Format | HTM / HTML PDF |
ISSN | 00219258 |
Journal | Journal of Biological Chemistry (JBC) |
Issue Number | 52 |
Volume Number | 276 |
DOI | 10.1074/jbc.M104346200 |
e-ISSN | 1083351X |
Language | English |
Publisher | American Society for Biochemistry and Molecular Biology |
Publisher Date | 2001-12-28 |
Access Restriction | Open |
Subject Keyword | Phospholipase C (PLC) Guanine nucleotide-binding protein (G protein) Myristic acid attachment signal G protein-coupled receptor kinase-C terminus (MAS-GRK-ct) Thyrotropin-releasing hormone (TRH) Regulator of G protein signaling (RGS) MECHANISMS OF SIGNAL TRANSDUCTION |
Content Type | Text |
Resource Type | Article |
Subject | Molecular Biology Cell Biology Biochemistry |
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