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Heat Shock Protein DnaK – Substrate of Actin-Specific Bacterial Protease ECP32

A. V. Morozova*, S. Yu. Khaitlina, and A. Yu. Malinin

Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, 194064 St. Petersburg, Russia; fax: (812) 297-0328; E-mail: avmoro@gmail.com

* To whom correspondence should be addressed.

Received September 15, 2010; Revision received December 20, 2010
It has been found that actin-specific bacterial protease ECP32 cleaves prokaryotic heat shock protein DnaK, which belongs to the family of heat shock proteins with molecular weight 70 kDa. We propose a new one-step method for DnaK purification using heat treatment. The technique yields ~1 mg of partially purified DnaK from 25 g of wet bacterial biomass. Polyclonal antibodies against DnaK were obtained. The degree of ECP32 catalyzed proteolysis of partially purified DnaK and that of DnaK in initial cell extracts was compared.
KEY WORDS: protease ECP32, DnaK, protein purification, limited proteolysis

DOI: 10.1134/S0006297911040080