Method for the cellular high-throughput-detection of receptor ligand interactions
OwnerAlbright E. Sippel, André Zimmermanninventors
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OwnerAlbright E. Sippel, André Zimmermanninventors
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The invention relates to cells which comprise a membrane receptor which comprises a ligand-binding section, a membrane-localization signal and a mediator section, and which is characterized in that only when there is binding or, alternatively, only when there is a lack of binding of a ligand to the ligand-binding section of the membrane receptor is a structural change brought about with effects on the mediator section to result in binding of an effector protein or polypeptide, which is capable of activating a Ras or Ras-like signal pathway in the cell, to the component of the membrane, where appropriate via other proteins or polypeptides (adaptors). It further relates to assay methods employing these cells, which are used, inter alia, to detect specific interactions between said membrane receptor and a ligand, and kits for use in these assays.
An isolated transformed yeast cell comprising, a human epidermal growth factor (EGF) membrane receptor and a fusion protein comprising an effector polypeptide which is a constitutively active human ras polypeptide fused to an adaptor polypeptide which is Grb2 polypeptide, wherein upon binding of ligand to said receptor, said fusion protein binds to said receptor via said adaptor polypeptide, and wherein said fusion protein is capable of activating a Ras-signaling pathway in said cell upon binding of ligand to said EGF receptor.
Dependencies are inferred from the claim text.