2. (10 points). Describe how fluorescence polarization (or…
2. (10 points). Describe how fluorescence polarization (or anisotropy) measurements can be used to measure a macromolecule/ligand interaction. Assume the small molecule ligand can emit fluorescence in this example. a. (4 points). What property of the light does one take advantage of to assess the binding reaction? Describe how this property leads to the ability to distinguish free from bound ligand. b. (3 points). What property of the fluorophore should one consider when designing these experiments (and why)? c. (3 points). In the case that the ligand is not fluorescent, one might chemically attach a fluorophore to the ligand. What would happen to the measured polarization if the fluorophore was linked to the ligand by a relatively long linker compared to very short linker?