... ofburied hydrophobic residues, we studied the bindingof the fluorescent probe, 1,8-anilinonaphtalene sulfonicacid (ANS), to hydrophobic side chains in proteins:this binding results in a marked increase ... for temperature- and urea-induced unfolding as determined by different approaches on thewild-type and mutant proteins of D- amino acid oxidase (DAAO).Method Wild typea,bDLOOPa,bW243I W243YTemperature ... position during flavin bindingFig. 4. (A) Time course of protein fluorescence change at 340 nmduring the binding of FAD to wild-type (d) and W243Y (n) apopro-teins at 15 °C and pH 7.5. Proteins were...