FLUORESCENCE SPECTROSCOPY AS A TOOL TO INVESTIGATE PROTEIN INTERACTIONS

Recent advances in the use of fluorescence spectroscopy to study prote in interactions have primarily involved combinations of classic fluore scence techniques, novel probe and coupling chemistries, and advances in laser excitation and detection capabilities. For example, new coupl ing strategies for fluorescent probes have allowed the first determina tion of the Delta G degrees describing the insertion of a protein into a membrane. Fluorescently labeled oligonucleotides with specific prot ein-binding sequences have been used to study both protein-DNA associa tions and oligonucleotide hybridization using anisotropy changes. The first kinetic data describing a DNA-protein binding event was collecte d with stopped-flow fluorescence instrumentation. Combining scanning f luctuation correlation spectroscopy with a two-photon excitation sourc e improved this technique so that it may now be used to study protein self-associations.