title: Chemical modification of tryptophan residues and stability changes in proteins.
abstract: The role of tryptophan residues in the stability of proteins was studied by ozone oxidation, which causes a small change in the tryptophan side chain. Trp 187 of the constant fragment of a type lambda immunoglobulin light chain, Trp 59 of ribonuclease T1, and Trp 62 of hen egg white lysozyme were oxidized specifically by ozone to N'-formylkynurenine or kynurenine. Judging from their circular dichroic and fluorescence spectra, these modified proteins were found to be the same as those of the respective intact proteins. However, even the slight modification of a single tryptophan residue produced a large decrease in the stability of these proteins to guanidine hydrochloride and heat. The smaller the extent of exposure of the tryptophan residue, the greater the effect of the modification on the stability. The formal kinetic mechanism of unfolding and refolding by guanidine hydrochloride of the CL fragment was not altered by tryptophan oxidation, but the rate constants for unfolding and refolding changed. The thermal unfolding transitions were analyzed to obtain the thermodynamic parameters. The enthalpy and entropy changes for the modified proteins were larger than the respective values for the intact proteins.
authors: Department of Biology, Faculty of Science, Osaka University, Japan.
authors: T
authors: T
authors: Okajima
authors: Y
authors: Y
authors: Kawata
authors: K
authors: K
authors: Hamaguchi
mesh_terms: D000818:Animals; D002645:Chickens; D002942:Circular Dichroism; D004529:Egg White; D004795:Enzyme Stability; D019791:Guanidine; D006146:Guanidines; D006358:Hot Temperature; D007136:Immunoglobulins / Q000737:chemistry; D007700:Kinetics; D009113:Muramidase / Q000737:chemistry*; D010126:Ozone / Q000378:metabolism; D011487:Protein Conformation; D006163:Ribonuclease T1 / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D014364:Tryptophan / Q000737:chemistry*
chemical_list: D006146:Guanidines; D007136:Immunoglobulins; D010126:Ozone; D014364:Tryptophan; D006163:Ribonuclease T1; D009113:Muramidase; D019791:Guanidine