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Formation and quantification of protein complexes between peroxisomal alcohol oxidase and GroEL
Author(s) -
Evers Melchior E.,
Langer Thomas,
Harder Wim,
Hartl Franz-Ulrich,
Veenhuis Marten
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)80653-x
Subject(s) - groel , chemistry , biochemistry , polyacrylamide gel electrophoresis , gel electrophoresis , chromatography , enzyme , escherichia coli , gene
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.