Mutations Leu427, Asn428, and Leu431 residues within transmembrane domain-I-segment 6 attenuate ginsenoside-mediated L-type Ca(2+) channel current inhibitions.
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.
Choi
Sun-Hye
SH
Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University.
Lee
Jun-Ho
JH
Pyo
Mi Kyung
MK
Lee
Byung-Hwan
BH
Shin
Tae-Joon
TJ
Hwang
Sung-Hee
SH
Kim
Bo-Ra
BR
Lee
Sang-Mok
SM
Oh
Jae-Wook
JW
Kim
Hyoung-Chun
HC
Bae
Chun Sik
CS
Rhim
Hyewhon
H
Nah
Seung-Yeol
SY
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
D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D036145:Ginsenosides; C097367:ginsenoside Rg3; D001216:Asparagine; D007930:Leucine