Received September 4, 1997
The advantages and some limitations of the use of fluorescence methods for the quantitative determination of the thermodynamics of protein unfolding transitions (i.e., induced by temperature or chemical denaturant) are discussed. Advantages include the sensitivity, multi-dimensional nature of the data, wide amenable concentration range, high signal-to-noise, rapidity of measurement, and adaptability to a variety of sample compartments. Aside from the need for a probe, some problems associated with the method involve the handling of baselines for the pre- and post-transition regions and the difficulty (shared by most other methods) of discerning whether the transition is two-state or multi-state.
KEY WORDS: fluorescence of proteins, protein unfolding, thermodynamics of protein unfolding, tryptophan fluorescence in proteins