title: Mutations Leu427, Asn428, and Leu431 residues within transmembrane domain-I-segment 6 attenuate ginsenoside-mediated L-type Ca(2+) channel current inhibitions.
abstract: Many lines of evidences have shown that Panax ginseng exhibits beneficial effects on cardiovascular systems. We previously demonstrated that ginsenoside Rg(3) (Rg(3)), one of active ingredients of Panax ginseng, inhibits Ca(2+) channel currents in a stereospecific manner and affects the steady-state activation but not inactivation. This points a possibility that Rg(3) regulates Ca(2+) channels through specific interaction site(s) for Ca(2+) influx inhibition through Ca(2+) channels. However, it was not known how Rg(3) interacts with Ca(2+) channel proteins. In the current study, we sought to identify these site(s) in Xenopus oocytes expressing cardiac wild-type and mutant L(alpha(1C))-type Ca(2+) channels using the two-microelectrode voltage-clamp technique. To this end, we assessed how various point mutations of the L-type Ca(2+) channel affected the Rg(3) action. Mutations of L427R, N428R and L431K in transmembrane domain-I-segment 6 (IS6) of the channel significantly attenuated the Rg(3) action and caused rightward shifts in dose-response curves. Rg(3) treatment produced a negative shift in the inactivation voltage but did not alter the steady-state activation voltage, and none of the mutant channels affected the Rg(3)-induced negative shift of inactivation voltage. Rg(3) had no effects on inactivation time constant in wild-type and mutant channels. These results indicate that Rg(3) inhibition of L-type Ca(2+) channel currents is attenuated by mutations of Leu427, Asn428 and Leu431 in transmembrane IS6 residues. Leu427, Asn428 and Leu431 residues of the L-type Ca(2+) channel play important roles in the Rg(3) effect on channel properties.
authors: Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University.
authors: Sun-Hye
authors: SH
authors: Choi
authors: Jun-Ho
authors: JH
authors: Lee
authors: Mi Kyung
authors: MK
authors: Pyo
authors: Byung-Hwan
authors: BH
authors: Lee
authors: Tae-Joon
authors: TJ
authors: Shin
authors: Sung-Hee
authors: SH
authors: Hwang
authors: Bo-Ra
authors: BR
authors: Kim
authors: Sang-Mok
authors: SM
authors: Lee
authors: Jae-Wook
authors: JW
authors: Oh
authors: Hyoung-Chun
authors: HC
authors: Kim
authors: Chun Sik
authors: CS
authors: Bae
authors: Hyewhon
authors: H
authors: Rhim
authors: Seung-Yeol
authors: SY
authors: Nah
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D001216:Asparagine / Q000235:genetics*; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D020746:Calcium Channels, L-Type / Q000096:biosynthesis / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D005260:Female; D036145:Ginsenosides / Q000494:pharmacology*; D015640:Ion Channel Gating / Q000187:drug effects; D007930:Leucine / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D014982:Xenopus laevis
chemical_list: D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D036145:Ginsenosides; C097367:ginsenoside Rg3; D001216:Asparagine; D007930:Leucine