title: Relationship between the wavelength maximum of a protein and the temperature dependence of its intrinsic tryptophan fluorescence intensity.
abstract: Proper determination of the temperature dependence of intrinsic tryptophan fluorescence intensity in native and denatured states is an essential prerequisite for extracting the free energy of protein unfolding from the thermal denaturation profile. The most common method employed in determining the temperature dependence of these conformations is through the determination of slopes of pre- and post-transition baselines. However, simulations of protein unfolding profiles suggest that this method does not work for marginally stable proteins. We show herein that the temperature dependence of the fluorescence intensity of N-acetyl tryptophanamide (NATA) in organic solvents and water may be used to represent the temperature dependence of the fluorescence intensity of tryptophan in native and denatured conformations of a protein, respectively. The wavelength of the emission maximum, lambda (max), of N-acetyl tryptophanamide (NATA) in a particular solvent or tryptophan in proteins is related to the temperature dependence (m) of its fluorescence intensity by the equation: m (K(-1)) = (-0.000299 +/- 2.2 x 10(-5) K(-1) nm(-1)) x lambda (max) (nm) + (0.0919 +/- 0.0025 K(-1)).
authors.affiliation: Department of Chemistry, Indian Institute of Technology, Delhi, India.
authors.forename: Komal
authors.initials: K
authors.lastname: Saini
authors.forename: Shashank
authors.initials: S
authors.lastname: Deep
mesh_terms: D003198:Computer Simulation; D005453:Fluorescence; D007700:Kinetics; D008956:Models, Chemical; D009010:Monte Carlo Method; D009930:Organic Chemicals / Q000737:chemistry; D011489:Protein Denaturation; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013696:Temperature; D014364:Tryptophan / Q000031:analogs & derivatives / Q000737:chemistry*; D014867:Water / Q000737:chemistry
chemical_list: D009930:Organic Chemicals; D011506:Proteins; D012997:Solvents; D014867:Water; C016952:N-acetyltryptophanamide; D014364:Tryptophan