Lysozyme dimer formation on lysozyme oxidation with B./2 as studied by fluorescence evolution

Lysozyme dimers produced on oxidation of lysozyme with Br2anion radicals in aqueous solutions exhibit a fluorescence spectrum (lambdasub(max) = 400 nm) closely similar to that of bi-tyrosine. This suggests that the dimer is likely to have a tyrosine-tyrosine bond resulting from the combination of tyrosine phenoxy radicals of two lysozyme molecules. Kinetic studies on dimer formation were made by measuring time-dependent fluorescence after pulsed-electron irradiation over wide pH range. The results lead to the following conclusions. The second-order growth of the dimer fluorescence observed at pH 10.7-12.5 reflects the combination process of the lysozyme radicals, which is rate-determining in the pH range. On the other hand, the first-order growth observed at pH 6.8-10.2 is attributable to the enolization of the keto-form of the dimer. A tentative reaction scheme is proposed for the dimer formation. (author)