Me. Evers et al., AFFINITY PURIFICATION OF MOLECULAR CHAPERONES OF THE YEAST HANSENULA-POLYMORPHA USING IMMOBILIZED DENATURED ALCOHOL OXIDASE, FEBS letters, 321(1), 1993, pp. 32-36
We used peroxisomal alcohol oxidase (AO) for the affinity purification
Of molecular chaperones from yeasts. Methodical studies showed that u
p to 0.8 mg of purified bacterial GroEL was able to bind per ml of imm
obilized denatured AO column material. Using crude extracts of Hansenu
la polymorpha or Saccharomyces cerevisiae, several proteins were speci
fically eluted with Mg-ATP which were recognized by antibodies against
hsp60 or hsp70. One H. polymorpha 70 kDa protein was strongly induced
during growth at elevated temperatures, whereas a second 70 kDa prote
in as well as a 60 kDa protein showed strong protein sequence homology
to mitochondrial SSC1 and hsp60, respectively, from S. cerevisiae.