Formation and quantification of protein complexes between peroxisomal alcohol oxidase and GroEL

FEBS Letters

Harder, Wim¹  Hartl, Franz-Ulrich²  Evers, Melchior E.¹  Langer, Thomas³  Veenhuis, Marten¹ 

[1] Laboratory for Electron Microscopy and Department of Microbiology, Biological Centre, University of Groningen, Kerklaan 30, 9751 NN Haren, The Netherlands;Program in Cellular Biochemistry and Biophysics, Rockefeller Research Laboratory, Sloane Kettering Institute, Box 520, 1275 York Avenue, New York, NY 10021, USA;Institut für Physiologische Chemie, Universität München, Goethestrasse 33, D-8000 München 2, Germany

关键词: Alcohol oxidase;    GroEL;    HSP;    Non-denaturing gel electrophoresis;    Peroxisomal matrix protein;    Protein complex formation;   

DOI  :  10.1016/0014-5793(92)80653-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have studied the use of yeast peroxisomal alcohol oxidase (AO) as a model protein for in vitro binding by GroEL. Dilution of denatured AO in neutral buffer leads to aggregation of the protein, which is prevented by the addition or GroEL. Formation of complexes between GroEL and denatured AO was demonstrated by a gel-shift assay using non-denaturing polyacrylamide gel electrophoresis, and quantified by laser-densitometry of the gels. In the presence of MgAMP-PNP or MgADP the affinity of GroEL for AO was enhanced. Under these conditions up to 70% of the purified GroEL formed a complex with this protein. Release was stimulated at room temperature by MgATP, and was further enhanced by addition or GroFS.

【 授权许可】

Unknown