METHODS FOR DETECTING CHANGES TO CELLULAR COMPONENTS

摘要

A method of detecting the state, or the change in state, of a macromolecular component of a cell which can exist in a first state or a second state wherein in its first state the macromolecular component is substantially incapable of being bound by a sensor molecule and in its second state the macromolecular component is able to be bound by the sensor molecule, the method comprising the steps of (1) providing a sample comprising the macromolecular component wherein the macromolecular component is bound to a donor fluorophore; (2) contacting the sample provided in (1) with a sensor molecule which is able to bind the macromolecular component in its second state but is substantially incapable of binding the macromolecular component in its first state wherein the sensor molecule is bound to an acceptor molecule; and (3) detecting fluorescence resonance energy transfer (FRET) between the donor fluorophore bound to the macromolecular component and the acceptor molecule bound to the sensor molecule wherein FRET is measured by the decrease in the fluorescence lifetime of the donor fluorophore.