GLYCEROL-INDUCED DEVELOPMENT OF CATALYTICALLY ACTIVE CONFORMATION OF CROTALUS-ADAMANTEUS L-AMINO-ACID OXIDASE IN-VITRO

The reconstitutable apoprotein of Crotalus adamanteus L-amino acid oxi dase was prepared using hydrophobic interaction chromatography. After reconstitution with flavin adenine dinucleotide, the resulting protein was inactive, with a perturbed conformation of the flavin binding sit e. Subsequently, a series of cosolvent-dependent compact intermediates was identified. The nearly complete activation of the reconstituted a poprotein and the restoration of its native flavin binding site was ac hieved in the presence of 50% glycerol. We provide evidence that in ad dition to a merely stabilizing effect of glycerol on native proteins, glycerol can also have a restorative effect on their compact equilibri um intermediates, and we suggest the hydrophobic effect as a dominatin g force in this in vitro-assisted restorative process.