[논문]Effect of Calmodulin on Ginseng Saponin-Induced <tex>$Ca^{2+}$</tex>-Activated <tex>$Cl^{-}$</tex> Channel Activation in Xenopus laevis Oocytes

Lee Jun-Ho; Jeong Sang-Min; Lee Byung-Hwan; Kim Jong-Hoon; Ko Sung-Ryong; Kim Seung-Hwan; Lee Sang-Mok; Nah Seung-Yeol

doi:10.1007/bf02977670

Effect of Calmodulin on Ginseng Saponin-Induced $Ca^{2+}$-Activated $Cl^{-}$ Channel Activation in Xenopus laevis Oocytes 원문보기

Archives of pharmacal research : a publication of the Pharmaceutical Society of Korea, v.28 no.4, 2005년, pp.413 - 420

Lee Jun-Ho (Research Laboratory for the Study of Ginseng Signal Transduction and Department of Physiology, College of Veterinary Medicine, Konkuk University) , Jeong Sang-Min (Research Laboratory for the Study of Ginseng Signal Transduction and Department of Physiology, College of Veterinary Medicine, Konkuk University) , Lee Byung-Hwan (Research Laboratory for the Study of Ginseng Signal Transduction and Department of Physiology, College of Veterinary Medicine, Konkuk University) , Kim Jong-Hoon (Research Laboratory for the Study of Ginseng Signal Transduction and Department of Physiology, College of Veterinary Medicine, Konkuk University) , Ko Sung-Ryong (KT & G Central Research Institute) , Kim Seung-Hwan (Department of Exercise Science, College of Natural Science, Chungbuk National University) , Lee Sang-Mok (Research Laboratory for the Study of Ginseng Signal Transduction and Department of Physiology, College of Veterinary Medicine, Konkuk University) , Nah Seung-Yeol (Research Laboratory for the Study of Ginseng Signal Transduction and Department of Physiology, College of Veterinary Medicine, Konkuk Univ)

Abstract ▼ AI-Helper

We previously demonstrated the ability of ginseng saponins (active ingredients of Panax ginseng) to enhance $Ca^{2+}$-activated $Cl^{-}$ current. The mechanism for this ginseng saponin-induced enhancement was proposed to be the release of $Ca^{2+}$ from $IP_{3}-sensitive$ intracellular stores through the activation of PTX-insensitive $G\alpha_{q/11}$ proteins and PLC pathway. Recent studies have shown that calmodulin (CaM) regulates $IP_{3}$ receptor-mediated $Ca^{2+}$ release in both $Ca^{2+}-dependent$ and -independent manner. In the present study, we have investigated the effects of CaM on ginseng saponin-induced $Ca^{2+}$-activated $Cl^{-}$ current responses in Xenopus oocytes. Intraoocyte injection of CaM inhibited ginseng saponin-induced $Ca^{2+}$-activated $Cl^{-}$ current enhancement, whereas co-injection of calmidazolium, a CaM antagonist, with CaM blocked CaM action. The inhibitory effect of CaM on ginseng saponin-induced $Ca^{2+}$-activated $Cl^{-}$ current enhancement was dose- and time-dependent, with an $IC_{50} of 14.9\pm3.5 {\mu}M$. The inhibitory effect of CaM on saponin's activity was maximal after 6 h of intraoocyte injection of CaM, and after 48 h the activity of saponin recovered to control level. The half-recovery time was calculated to be $16.7\pm4.3 h$. Intraoocyte injection of CaM inhibited $Ca^{2+}$-induced $Ca^{2+}$-activated $Cl^{-}$ current enhancement and also attenuated $IP_{3}$-induced $Ca^{2+}$-activated $Cl^{-}$ current enhancement. $Ca^{2+}$/CaM kinase II inhibitor did not inhibit CaM-caused attenuation of ginseng saponin-induced $Ca^{2+}$-activated $Cl^{-}$ current enhancement. These results suggest that CaM regulates ginseng saponin effect on $Ca^{2+}$-activated $Cl^{-}$ current enhancement via $Ca^{2+}$-independent manner.

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가설 설정

1. Inset, peak outward currents (meant S.E.M; n=14-16 each) recorded in oocytes injected with H₂O or 40 M CaM in the presence of GTS (50 p.g/mL). The curve described by th으 sd line was fit by the Hill equation as described in Method sections.

대상 데이터

The ginseng total saponins (GTS) were a kind gift from Korea Ginseng Cooperation (Taejon, Korea). GTS contained Rb〔 (17.1%), Rb2 (9.07%), Rc (9.65%), Rd (8.26%), Re (9%), Rf (3%), Rg1 (6.4%), Rg2 (4.2%), Rg3 (3.8%), Ro (3.8%), Ra (2.91%) and other minor ginsenosides. GTS was dil니ted with bath medium, ND96 before use.
A custom-made Plexiglas net chamber was used for two-electrode voltage-clamp recordings. The chamber was constructed by milling two concentric wells into the chamber bottom (diameter/height: upper well: 8/3 mm, lower well: 6/5 mm) and gluing plastic meshes (-0.4-mm grid diameter) onto the bottom of the upper well. The perf니sion inlet (~1-mm diameter) was formed through the wall of the lower well, and the suction tube was placed on the edge of the upper well.

성능/효과

Second, we showed that intraoocyte injected CaM blocked ginseng saponin-induced Ca2+-activated Cl~ current response in a concentration-dependent manner. Third, the inhibitory effect of CaM on ginseng saponin-induced Ca2+-activated Cl~ current enhancement was persistent but reversed slowly, since ginseng saponin effect on Ca2+-activated Cl- current enhancement completely recovered 48 h after intraoocyte injection of CaM. Thus, the half-recovery time for ginseng saponin effect on Ca?七activated Cl~ channel was about 17 h, suggesting that the binding affinity of CaM to IP3 receptor or other regulatory site(s) might be high.

참고문헌 (26)

Adkins, C. E., Morris, S. A., De Smedt, H., Sienaert, I., Torok, K., and Taylor, C. W., $Ca^{2+}$ -calmodulin inhibit $Ca^{2+}$ release mediated by type-1, -2, and -3 inositol trisphosphate receptors. Biochem. J., 345, 357-363 (2000)
Berridge, M. J. and Irvine, R. F., Inositol triphosphates and cell signalling. Nature, 341, 197-205 (1989)

상세보기
Berridge, M. J., Bootman, M. D., and Lipp, P., Calcium - a life and death signal. Nature (Lond.), 395, 645-648 (1998)

상세보기
Cardy, T. J. A. and Taylor, C. W., A novel role for calmodulin: $Ca^{2+}$ -independent inhibition of type-1 inositol trisphosphate receptors. Biochem. J., 334, 447-455 (1998)

상세보기
Choi, S., Rho, S. H., Jung, S. Y., Kim, S. C., Park, C. S., and Nah, S. Y., A novel activation of $Ca^{2+}$ -activated $Cl^{-}$ channel in Xenopus oocytes by ginseng saponins: evidence for the involvement of phospholipase C and intracellular $Ca^{2+}$ mobilization. Br. J. Pharmacol., 132, 641-648 (2001a)

상세보기
Choi, S., Kim, H. J., Ko, Y. S., Jeong, S. W., Kim, Y. I., Simonds, W. F., Oh, J. W., and Nah, S. Y., $G\alpha_{q/11}$ coupled to mammalian phospholipase C b3-like enzyme mediates the ginsenoside effect on $Ca^{2+}$ -activated $Cl^-$ current in the Xenopus oocyte. J. Biol. Chem., 276, 48797-48802 (2001b)

상세보기
Dascal, N., Yekuel, R., and Oron, Y., Acetylcholine promotes progesterone-induced maturation of Xenopus oocytes. J. Exp. Zool., 230, 131-135 (1984)

상세보기
Fukunaga, K., Miyamoto, E., and Soderling T. R., Regulation of $Ca^{2+/}$ calmodulin-dependent protein kinase II by brain gangliosides. J. Neurochem., 54, 103-109 (1990)

상세보기
Gnegy, M. E., Calmodulin in neurotransmitter and hormone action. Ann. Rev. Pharmacol. Toxicol., 33, 45-70 (1993)

상세보기
Hartzell, H. C., Activation of different $Cl^-$ currents in Xenopus oocytes by Ca liberation from stores and by capacitative Ca Influx. J. Gen. Physiol., 108, 157-175 (1996)

상세보기
Jeong, S. M., Lee, J. H., Kim, S., Rhim, H., Lee B. H., Kim, J. H., Oh, J. W., and Lee, S. M., Ginseng saponins induce store-operated calcium entry in Xenopus oocytes. Br. J. Pharmacol., 142, 585-593 (2004)

상세보기
Kasri, N. N., Bultynck, G., Sienaert, I., Callewaert, G., Erneux, C., Missiaen, L., Pary, J. B., and De Smedt, H., The role of calmodulin for inositol 1,4,5-trisphosphate receptor function. Biochim. Biophys. Acta, 1600, 19-31 (2002)

상세보기
Kuruma, A. and Hartzell, H. C., Dynamics of calcium regulation of chloride currents in Xenopus oocytes. Am. J. Physiol., 276, C161-C175 (1999)

상세보기
Lechleiter, J. D. and Clapham, D. E., Molecular mechanisms of intracellular calcium excitability in Xenopus laevis oocytes. Cell, 69, 283-294 (1992)

상세보기
Lee, J. H., Jeong, S. M., Lee, B. H., Kim, D. H., Kim, J. H., Kim, J. I., and Nah, S. Y., Prevention of ginsenoside-induced desensitization of $Ca^{2+}$ -activated $Cl^-$ currents by microinjection of inositol hexakisphosphate ( $InsP_6$ ) in Xenopus laevis oocytes: involvement of GRK2 and $\beta-arrestin$ I. J. Biol. Chem., 279, 9912-9921 (2004)

상세보기
Liu, M., Chen, T. Y., Ahamed, B., Li, J., and Yau, K. W., Calciumcalmodulin modulation of the olfactory cyclic nucleotide gated cation channel. Science, 266, 1348-1354 (1994)

상세보기
Matifat, F., Fournier, F., Lorca, T., Capony, J. P., Brule, G.., and Collin, T., Involvement of the $Ca^{2+}$ /calmodulin protein kinase II pathway in the $Ca^{2+}$ -mediated regulation of the capacitative $Ca^{2+}$ entry in Xenopus oocytes. Biochem. J., 322, 267-272 (1997)

상세보기
Missiaen, L., Parys, J. B., Weidema, A. F., Sipma, H., Valingen, S., De Smet, P., Callewaert, G., and De Smedt, H., The bellshaped $Ca^{2+}$ dependence of the inositol 1,4,5-trisphosphateinduced $Ca^{2+}$ release is modulated by $Ca^{2+}$ /calmodulin. J. Biol. Chem., 274, 13748-13751 (1999)

상세보기
Nah, S. Y., Ginseng, recent advances and trend. Korea J. Ginseng Sci., 21, 1-12 (1997)
Parekh A. B., Interaction between capacitative $Ca^{2+}$ influx and $Ca^{2+}$ -dependent $Cl^-$ currents in Xenopus oocytes. Pflugers Arch-Eur. J. Physiol., 430, 954-963 (1995)

상세보기
Parekh, A. B. and Penner, R., Store depletion and calcium influx. Physiol. Rev., 77, 901-930 (1997)

상세보기
Parys, J. B., Sernett, S. W., DeLisle, S., Snyder, P. M., Welsh, M. J., and Campbell, K. P., Isolation, characterization, and localization of the inositol 1,4,5-triphosphate receptor protein in Xenopus laevis oocytes. J. Biol. Chem., 267, 18776-18782 (1992)
Sienaert, I., Kasri, N. N., Vanlingen, S. Parys, J. B., Callewaert, G., Messeian, L., and de Smedt, L., Localization and function of a calmodulin/apocalmodulin binding domain in the Nterminal part of the type-1 inositol 1,4,5-trisphosphate receptor. Biochem. J., 365, 269-277 (2002)

상세보기
Taylor, C. W. and Laude, A. J., $IP_3$ receptors and their regulation by calmodulin and cytosolic $Ca^{2+}$ . Cell Calcium, 32, 321-334 (2002)

상세보기
Yamada, M., Miyawaki, A., Saito, K., Nakajima, M., Yamamoto- Hino, Y., Ryo, T., Furuichi, T., and Mikoshiba, K., The calmodulin-binding domain in the mouse type 1 inositol 1,4,5- trisphosphate receptor. Biochem. J., 308, 83-88 (1995)

상세보기
Waxham, M. N. and Aronowski, J., $Ca^{2+}$ /calmodulin-dependent protein kinase II is phosphorylated by protein kinase C in vitro. Biochemistry, 32, 2923-2930 (1993)

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