q/11 Coupled to Mammalian Phospholipase C β3-like Enzyme Mediates the Ginsenoside Effect on Ca2+-activated Cl - Current in the Xenopus Oocyte

Seok Choi, Hyun Ju Kim, Yoo Seung Ko, Seong Woo Jeong, Yang In Kim, William F. Simonds, Jae Wook Oh, Seung Yeol Nah

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Recently we demonstrated that ginsenosides, the active ingredients of Panax ginseng, enhanced Ca2+-activated Cl- current in the Xenopus 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 Gα12 and GαoA cRNA injection had no significant effect. The changes following Gα q cRNA injection were prevented when Gβ1γ 2 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 Gα q cRNA injection. The effects of GαqQ209L cRNA injection, however, were not prevented by co-injection of Gβ 1γ2 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 Gα q/11 coupled to mammalian PLC β3-like enzyme mediates ginsenoside effect on Ca2+-activated Cl- current in the Xenopus oocyte.

Original languageEnglish
Pages (from-to)48797-48802
Number of pages6
JournalJournal of Biological Chemistry
Volume276
Issue number52
DOIs
Publication statusPublished - 2001 Dec 28

Fingerprint

Ginsenosides
Complementary RNA
Type C Phospholipases
Xenopus
Oocytes
Injections
Enzymes
GTP-Binding Proteins
Protein Subunits
Guanosine Triphosphate
Protein Isoforms
GTP-Binding Protein Regulators
Signal transduction
Panax
Antibodies
Pertussis Toxin
Protein C
Hydrolysis
Signal Transduction
Carrier Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

q/11 Coupled to Mammalian Phospholipase C β3-like Enzyme Mediates the Ginsenoside Effect on Ca2+-activated Cl - Current in the Xenopus Oocyte. / Choi, Seok; Kim, Hyun Ju; Ko, Yoo Seung; Jeong, Seong Woo; Kim, Yang In; Simonds, William F.; Oh, Jae Wook; Nah, Seung Yeol.

In: Journal of Biological Chemistry, Vol. 276, No. 52, 28.12.2001, p. 48797-48802.

Research output: Contribution to journalArticle

Choi, Seok ; Kim, Hyun Ju ; Ko, Yoo Seung ; Jeong, Seong Woo ; Kim, Yang In ; Simonds, William F. ; Oh, Jae Wook ; Nah, Seung Yeol. / q/11 Coupled to Mammalian Phospholipase C β3-like Enzyme Mediates the Ginsenoside Effect on Ca2+-activated Cl - Current in the Xenopus Oocyte. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 52. pp. 48797-48802.
@article{72ab8759b3854aab84c3d09e26c48605,
title = "Gαq/11 Coupled to Mammalian Phospholipase C β3-like Enzyme Mediates the Ginsenoside Effect on Ca2+-activated Cl - Current in the Xenopus Oocyte",
abstract = "Recently we demonstrated that ginsenosides, the active ingredients of Panax ginseng, enhanced Ca2+-activated Cl- current in the Xenopus 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 Gα12 and GαoA cRNA injection had no significant effect. The changes following Gα q cRNA injection were prevented when Gβ1γ 2 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 Gα q cRNA injection. The effects of GαqQ209L cRNA injection, however, were not prevented by co-injection of Gβ 1γ2 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 Gα q/11 coupled to mammalian PLC β3-like enzyme mediates ginsenoside effect on Ca2+-activated Cl- current in the Xenopus oocyte.",
author = "Seok Choi and Kim, {Hyun Ju} and Ko, {Yoo Seung} and Jeong, {Seong Woo} and Kim, {Yang In} and Simonds, {William F.} and Oh, {Jae Wook} and Nah, {Seung Yeol}",
year = "2001",
month = "12",
day = "28",
doi = "10.1074/jbc.M104346200",
language = "English",
volume = "276",
pages = "48797--48802",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "52",

}

TY - JOUR

T1 - Gαq/11 Coupled to Mammalian Phospholipase C β3-like Enzyme Mediates the Ginsenoside Effect on Ca2+-activated Cl - Current in the Xenopus Oocyte

AU - Choi, Seok

AU - Kim, Hyun Ju

AU - Ko, Yoo Seung

AU - Jeong, Seong Woo

AU - Kim, Yang In

AU - Simonds, William F.

AU - Oh, Jae Wook

AU - Nah, Seung Yeol

PY - 2001/12/28

Y1 - 2001/12/28

N2 - Recently we demonstrated that ginsenosides, the active ingredients of Panax ginseng, enhanced Ca2+-activated Cl- current in the Xenopus 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 Gα12 and GαoA cRNA injection had no significant effect. The changes following Gα q cRNA injection were prevented when Gβ1γ 2 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 Gα q cRNA injection. The effects of GαqQ209L cRNA injection, however, were not prevented by co-injection of Gβ 1γ2 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 Gα q/11 coupled to mammalian PLC β3-like enzyme mediates ginsenoside effect on Ca2+-activated Cl- current in the Xenopus oocyte.

AB - Recently we demonstrated that ginsenosides, the active ingredients of Panax ginseng, enhanced Ca2+-activated Cl- current in the Xenopus 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 Gα12 and GαoA cRNA injection had no significant effect. The changes following Gα q cRNA injection were prevented when Gβ1γ 2 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 Gα q cRNA injection. The effects of GαqQ209L cRNA injection, however, were not prevented by co-injection of Gβ 1γ2 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 Gα q/11 coupled to mammalian PLC β3-like enzyme mediates ginsenoside effect on Ca2+-activated Cl- current in the Xenopus oocyte.

UR - http://www.scopus.com/inward/record.url?scp=0035965997&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035965997&partnerID=8YFLogxK

U2 - 10.1074/jbc.M104346200

DO - 10.1074/jbc.M104346200

M3 - Article

C2 - 11673455

AN - SCOPUS:0035965997

VL - 276

SP - 48797

EP - 48802

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 52

ER -