Quercetin activates human Kv1.5 channels by a residue I502 in the S6 segment.
1. The aims of the present study were to investigate the pharmacological effects of quercetin on wild-type (WT) and mutant (I502A) human (h) Kv1.5 channel currents (I(kur)) and to identify whether mutation in the S6 segment is critical to activation of I(kur) by quercetin. 2. Experiments were performed on WT and site-directed mutant hKv1.5 channels, which were stably expressed in Xenopus oocytes using the two-microelectrode voltage-clamp technique. 3. Quercetin increased WT hKv1.5 channel current in a concentration-, voltage- and time-dependent manner, with an EC(50) of 37.8 micromol/L and a negative shift in the steady state activation and inactivation curves. Quercetin accelerated channel activation and inactivation, significantly decreasing activation and inactivation time constants. However, mutating the I502 residue to Ala abolished the activating effect of quercetin. Quercetin did not modify the activation and inactivation kinetics of I502A channels. As an anti-oxidant, tanshinone IIA (4 micromol/L) inhibited the H(2)O(2)-induced activation of WT hKv1.5 channels. In contrast, quercetin had no significant effect. 4. We conclude that: (i) quercetin preferentially binds to and increases the current amplitude of WT hKv1.5 channels; (ii) Ile502, an aliphatic and neutral amino acid residue residing in the S6 segment, is important in quercetin binding; and (iii) quercetin-induced changes in the properties of WT hKv1.5 channels may be foreign to its own anti-oxidant action.
Yang
Lan
L
Cardio-Electrophysiological Research Laboratory, Medical College, Wuhan University of Science and Technology, Wuhan, China.
Ma
Ji-Hua
JH
Zhang
Pei-Hua
PH
Zou
An-Ruo
AR
Tu
Dan-Na
DN
D045784:Abietanes; D000200:Action Potentials / Q000187:drug effects; D021542:Amino Acids, Neutral / Q000235:genetics; D000818:Animals; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D015640:Ion Channel Gating / Q000187:drug effects; D017136:Ion Transport / Q000187:drug effects; D051666:Kv1.5 Potassium Channel / Q000235:genetics* / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D010616:Phenanthrenes / Q000494:pharmacology; D011485:Protein Binding; D011794:Quercetin / Q000494:pharmacology*; D013997:Time Factors; D014982:Xenopus laevis
D045784:Abietanes; D021542:Amino Acids, Neutral; D051666:Kv1.5 Potassium Channel; D010616:Phenanthrenes; C021751:tanshinone; D011794:Quercetin; D006861:Hydrogen Peroxide