alpha subunit for identification of modulators
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United States of America Patent
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Oct 8, 2013
Grant Date -
Feb 16, 2012
app pub date -
Dec 21, 2009
filing date -
Dec 22, 2008
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Abstract
Reagents, methods and kits for screening for compounds that modulate the activity of voltage-gated sodium channels (NaV), such as human NaV1.5/SC-N5A/hH1 are described. The reagents, methods and kits are based on mutated NaV alpha subunit polyptides of SEQ ID NO:5 with mutations at positions 372, 898, 1419 and 1711 (the DEKA motif) and at positions 11485, 1486 and 1487 (the IFM motif) resulting in increased permeability for a group IIA divalent cation (Ca++) and decreased inactivation rate. The mutant polypeptide is used in a method and kit for determining whether a test compound modulates the channel activity, preferably using a chimeric polypeptide (chameleon polypeptide) comprising calmodulin, a calmodulin binding protein (M13), and two fluorescent agents.
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