SCREENING METHOD FOR ION CHANNEL MODULATORS USING MUTATED BKCA CHANNEL

摘要

The present disclosure relates to a doubly mutated BKCa channel construct and a novel cell-based screening system for ion channel modulators using the same. A doubly mutated BKCa channel-comprising cell-based system of the present invention, when compared to a control group, shows remarkably increased fluorescence caused by membrane depolarization, regardless of whether there is a separate increase of [Ca2+]i, and shows activity (movement in the negative direction of a G/V curve) that is further triggered by a known activator (for example, CTBIC). Therefore, the system of the present disclosure is capable of more effectively and accurately analyzing the activity of an ion channel, and thus can be effectively applied not only to the separation/identification of ion channel modulators but also to screening for a therapeutic agent for a condition, disease or disorder related to the modulation of a BKCa channel.

主权项

1. A method for screening ion channel modulators, comprising:
(a) treating a cell comprising a nucleotide sequence encoding an amino acid sequence in which amino acids at positions G733 and N736 in a wild type BKCa channel gene are mutated with a test material; and (b) analyzing the activity of the mutated BKCa channel in the cell, wherein the test material is determined to be an ion channel activator when the test material potentiates the activity of the mutated BKCa channel, and the test material is determined to be an ion channel inhibitor when the test material inhibits the activity of the mutated BKCa channel.