Methods of screening for TRPM4b modulators
AssigneeThe Queen's Medical Center
InventorReinhold Penner
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Abstract
The invention relates, in part, to methods useful in identifying molecules, that bind TRPM4b, which modulate TRPM4b ion channel activity, and/or which alter expression of TRPM4b within cells. The TRPM4b channels as described herein comprise TRPM4b polypeptides, which are in turn encoded by TRPM4b nucleic acids. The ion channels described herein are preferably formed in HEK-293 cells and comprise one or more novel TRPM4b polypeptides, which exhibit one or more of the unique TRPM4b properties described herein.
Claims34
A method for screening for a candidate bioactive agent capable of binding to TRPM4b, said method comprising:
- a) contacting a TRPM4b protein or an amino-terminal fragment thereof with said candidate agent;
- and
- b) determining the binding of said candidate agent to said TRPM4b protein or amino-terminal fragment thereof.
A method for screening a candidate bioactive agent comprising
- a) contacting a TRPM4b channel with the candidate agent, and
- b) detecting whether said agent modulates the monovalent cationic permeability of said TRPM4b channel.
A method for screening for a candidate bioactive agent capable of modulating monovalent cation permeability of a TRPM4b channel, said method comprising:
- a) providing a recombinant cell comprising a recombinant nucleic acid comprising nucleic acid encoding TRPM4b and an inducible promoter operably linked thereto which is capable of expressing said TRPM4b, and further comprising a monovalent cation indicator;
- b) inducing said recombinant cell to express said TRPM4b;
- c) contacting said recombinant cell with a monovalent cation and said candidate agent;
- and
- d) detecting the intracellular levels of said monovalent cation with said indicator.
A method for measuring monovalent cation permeability of a TRPM4b channel, said method comprising:
- a) providing a recombinant cell wherein said cell comprises a recombinant nucleic acid which expresses TRPM4b and further comprises a monovalent cation indicator;
- b) contacting said recombinant cell with a monovalent cation which selectively interacts with said indicator to generate a signal;
- and
- c) measuring the intracellular levels of said monovalent cation by detecting said indicator signal.
A method for screening for a candidate bioactive agent capable of modulating expression of a TRPM4b protein or fragment thereof comprising:
- a) providing a recombinant cell capable of expressing a recombinant nucleic acid encoding a TRPM4b protein;
- b) contacting said cell with said candidate agent;
- and
- c) determining the effect of said candidate agent on the expression of said recombinant nucleic acid.
Dependencies are inferred from the claim text.