Me. Evers et al., IN-VITRO DISSOCIATION AND RE-ASSEMBLY OF PEROXISOMAL ALCOHOL OXIDASESOF HANSENULA-POLYMORPHA AND PICHIA-PASTORIS, FEBS letters, 368(2), 1995, pp. 293-296
We have studied the in vitro inactivation/dissociation and subsequent
reactivation/re-assembly of peroxisomal alcohol oxidases (AO) from the
yeasts Hansenula polymorpha and Pichia pastoris. Both proteins are ho
mo-oligomers consisting of eight identical subunits, each containing o
ne FAD as the prosthetic group. They were both rapidly inactivated upo
n incubation in 80% glycerol, due to their dissociation into the const
ituting subunits, which however still contained FAD. Dilution of disso
ciated AO in neutral buffer lead to reactivation of the protein due to
AO re-assembly, as was demonstrated by non-denaturing PAGE. After use
of mixtures of purified AO from H, polymorpha and P. pastoris active
hybrid AO oligomers mere formed, When prior to dissociation FAD was ch
emically removed from AO, reactivation or re-assembly did not occur in
dependent of externally added FAD.