AFFINITY PURIFICATION OF MOLECULAR CHAPERONES OF THE YEAST HANSENULA-POLYMORPHA USING IMMOBILIZED DENATURED ALCOHOL OXIDASE

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.