Reactivation-based molecular interaction sensors
First Claim
1. A method for detecting an interaction of binding ensemble members, comprising the steps of:
- providing a responder complex comprising linked components;
a responder molecule, an inhibitor of the responder molecule, and a first member of a binding ensemble;
providing a reactivator complex comprising a reactivator molecule linked to a second member of the binding ensemble;
combining the responder complex and the reactivator complex, where upon the interaction of the first and the second binding ensemble member, the reactivator competitively or allosterically displaces the inhibitor from the responder molecule; and
detecting an activity of the responder molecule, thereby detecting the interaction of the first and the second binding ensemble members.
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Abstract
This invention provides methods and systems for linking the functional activation of a responder molecule to the interaction of two or more binding ensemble members of interest either in vitro or in vivo, and thereby producing a signal, phenotype, or other functional output in response to the interaction of the binding ensemble members. The systems comprise a responder, an inhibitor of the responder, and an inhibitor of the inhibitor, or “reactivator” of the responder, and interacting components. Two binding ensemble members are complexed to the other components of the system in such a way that interaction of the binding ensemble members, either directly or via additional ensemble members, causes a shift in the equilibrium of inhibitor binding from the responder to the reactivator, thereby functionally activating the responder.
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Citations
29 Claims
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1. A method for detecting an interaction of binding ensemble members, comprising the steps of:
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providing a responder complex comprising linked components;
a responder molecule, an inhibitor of the responder molecule, and a first member of a binding ensemble;providing a reactivator complex comprising a reactivator molecule linked to a second member of the binding ensemble; combining the responder complex and the reactivator complex, where upon the interaction of the first and the second binding ensemble member, the reactivator competitively or allosterically displaces the inhibitor from the responder molecule; and detecting an activity of the responder molecule, thereby detecting the interaction of the first and the second binding ensemble members. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29)
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Specification