Affinity purification of molecular chaperones of the yeast Hansenula polymorpha using immobilized denatured alcohol oxidase

FEBS Letters

Harder, Wim¹  Huhse, Bettina²  Kunau, Wolf H.²  Hartl, Franz-Ulrich³  Evers, Melchior E.¹  Veenhuis, Marten¹  Titorenko, Vladimir I.¹ 

[1] Laboratory for Electron Microscopy, Biological Centre, University of Groningen, Kerklaan 30, 9751 NN Haren, The Netherlands;Institute for Physiological Chemistry, Medical Faculty, Ruhr University Bochum, Bochum, Germany;Program in Cellular Biochemistry and Biophysics, Rockefeller Research Laboratory, Sloane Kettering Institute, Box 520, 1275 York Avenue, New York, NY 10021, USA

关键词: Alcohol oxidase;    Chaperone;    HSP;    Peroxisome;    Saccharomyces cerevisiae;    Hansenula polymorpha;    Complex formation;   

DOI  :  10.1016/0014-5793(93)80615-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We used peroxisomal alcohol oxidase (AO) for the affinity purification of molecular chaperones from yeasts. Methodical studies showed that up to 0.8 mg of purified bacterial GroEL was able to bind per ml of immobilized denatured AO column material. Using crude extracts of Hansenula polymorpha or Saccharomyces cerevisiae, several proteins were specifically 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 protein as well as a 60 kDa protein showed strong protein sequence homology to mitochondrial SSCI and hsp60, respectively, from S. cerevisiae.

【 授权许可】

Unknown