Screening methods and assays for use with transmembrane proteins, particularly GPCRs.

Abstract

This invention provides methods and tools that can be used for assays, screenings, and drug discovery and development attempts. [Solution] An arrangement and method for screening membrane proteins are provided. The arrangement comprises a first fusion protein containing a membrane protein present in a boundary layer such as the wall of a cell, liposome, or vesicle, fused to one member of a binding pair, and a second fusion protein containing an intracellular ligand of the membrane protein, fused to the other member of the binding pair, the binding pair can generate a detectable signal.

Claims

1. Two or more immunoglobulin monovariable domains, each capable of binding to a different epitope, portion, domain, or subunit of a heterotrimeric G protein complex; and A pair of members, comprising at least a first and a second bonded member, capable of generating a detectable signal. Includes, A fusion protein in which the two or more immunoglobulin single variable domains and the members of the binding pair are linked or fused to one another.

2. The fusion protein according to claim 1, wherein the two or more immunoglobulin monovariable domains and the members of the binding pair are linked or fused to one another via a linker or spacer.

3. The fusion protein according to claim 1 or 2, wherein at least one of the immunoglobulin single variable domains is capable of specifically binding to the G alpha subunit of a heterotrimeric G protein complex.

4. A fusion protein according to any one of claims 1 to 3, comprising an immunoglobulin monovariable domain CA4435 (SEQ ID NO: 1).

5. A fusion protein according to any one of claims 1 to 3, comprising an immunoglobulin monovariable domain CA4437 (SEQ ID NO: 2).

6. The fusion protein according to any one of claims 1 to 3, comprising immunoglobulin monovariable domain CA4435 (SEQ ID NO: 1) and immunoglobulin monovariable domain CA4437 (SEQ ID NO: 2).

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