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THE RATES OF PHOTOLYSIS OF THE FOUR INDIVIDUAL TRYPTOPHAN RESIDUES IN UV EXPOSED CALF γ‐II CRYSTALLIN
Author(s) -
Tallmadge D. H.,
Borkman R. F.
Publication year - 1990
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1990.tb01723.x
Subject(s) - tryptophan , crystallin , chemistry , fluorescence , peptide , dithiothreitol , chromatography , irradiation , biochemistry , amino acid , physics , quantum mechanics , nuclear physics , enzyme
— Aqueous buffer solutions of the lens protein bovine γ‐II crystallin were irradiated at 295 nm in the presence of dithiothreitol to determine the individual photolysis susceptibilities of the four tryptophan residues. Reverse‐phase high performance liquid chromatography was utilized to compare the tryptic peptide maps before and after irradiation. Sequence analysis of collected tryptic peptides showed that the four tryptophans in calf γ‐II crystallin, TRP‐42, TRP‐68, TRP‐131, and TRP‐157 appeared in four distinct tryptic peptides. Fluorescence and absorption (diode array) monitoring of the eluting peptides allowed assessment of the changes in peptide absorbance and fluorescence following irradiation. Tryptophan fluorescence losses of (40 ± 15)%, (17 ± 4)%, (35 ± 5)% and (15 ± 4)% were observed for the peptides containing TRP‐42, TRP‐68. TRP‐131 and TRP‐157, respectively. Thus the four tryptophans in calf γ‐II crystallin did not all photolyze at the same rate. The rate differences are presumably related to the microenvironments of the individual tryptophan residues, and this is discussed in terms of the known crystal structure of calf γ‐II crystallin.