METHODS, REAGENTS AND KITS FOR THE IDENTIFICATION OF MODULATORS OF VOLTAGE-GATED SODIUM CHANNELS

摘要

Reagents, methods and kitsfor screening for compounds that modulatethe activity of voltage-gated sodium channels(NaV), such as human NaV1.5/SCN5A/hH1are described. The reagents,methods and kits are based on mutated NaValpha subunit polyptides of SEQ ID NO:5with mutations at positions 372, 898, 1419and 1711 (the DEKA motif) and at positions11485, 1486 and 1487 (the IFM motif)resulting in increased permeability for agroup IIA divalent cation (Ca++) and decreasedinactivation rate. The mutantpolypeptide is used in a method and kit fordetermining whether a test compound modulatesthe channel activity, preferably usinga chimeric polypeptide (chameleonpolypeptide) comprising calmodulin, acalmodulin binding protein (M13), and twofluorescent agents.