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Progressive anisotropy changes of the 1 μM HA-FRET probe in the presence of HA-ase. The observation was at the donor fluorescence. No changes in the fluorescence anisotropy were observed in the absence of the HA-ase. Two effects resulting from the HA-FRET cleavage are responsible for the anisotropy decrease: 1) smaller fragments of the macromolecule rotate faster, 2) a lifetime of fluorescein donor increases in the absence of acceptors.
