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Bio Protoc. 2017 Oct 5;7(19). pii: e2563. doi: 10.21769/BioProtoc.2563.

Protein Expression and Purification of the Hsp90-Cdc37-Cdk4 Kinase Complex from Saccharomyces cerevisiae.

Author information

1
Howard Hughes Medical Institute (HHMI) and the Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA 94158, USA.

Abstract

Interactions between Hsp90, its co-chaperone Cdc37 and kinases have been biochemically studied for over three decades and have been shown to be functionally important in organisms from yeast to humans. However, formation of a stable complex for structural studies has been elusive. In this protocol we describe expression and purification of Hsp90-Cdc37-Cdk4 kinase protein complex from Saccharomyces cerevisiae utilizing the viral 2A sequences to titrate the three proteins at similar levels.

KEYWORDS:

2A peptides; Cdc37; Cdk4; Chaperone; Hsp90; Kinase; Yeast protein expression

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