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

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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

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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.

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KEY WORDS: protease ECP32, DnaK, protein purification, limited proteolysis

DOI: 10.1134/S0006297911040080

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