    1.4.3.3 | holo- and apoprotein samples of hDAAO are incubated for 60 min at 15°C in a buffer containing different concentrations of urea, 0-6 M, and then refolded by 10fold dilution in 50 mM sodium diphosphate, pH 8.0, 5% glycerol and 15°C, in the presence of a 10fold molar excess of FAD. Chemical denaturation of hDAAO holoenzyme is partially reversible, 50% of the initial activity is recovered starting with the refolding from 4 M urea-denatured holoprotein, while the refolding of apoprotein is largely irreversible even at 2 M urea , 15-20% of recovery of enzymatic activity versus 90-100% for the holoenzyme and even in the presence of the cofactor |
701111 |
