DnaK ATPase-BD E.Coli
Description
Recombinant DnaK Substrate Binding Domain produced in E.Coli is a single, non-glycosylated polypeptide chain containing 384 amino acids and having a molecular mass of 41.6 kDa.
Product Specs
MGKIIGIDLG TTNSCVAIMD GTTPRVLENA EGDRTTPSII AYTQDGETLV GQPAKRQAVTNPQNTLFAIK RLIGRRFQDE EVQRDVSIMP FKIIAADNGD AWVEVKGQKM APPQISAEVLKKMKKTAEDY LGEPVTEAVI TVPAYFNDAQ RQATKDAGRI AGLEVKRIIN EPTAAALAYGLDKGTGNRTI AVYDLGGGTF DISIIEIDEV DGEKTFEVLA TNGDTHLGGE DFDSRLINYLVEEFKKDQGI DLRNDPLAMQ RLKEAAEKAK IELSSAQQTD VNLPYITADA TGPKHMNIKV TRAKLESLVE DLVNRSIEPL KVALQDAGLS VSDIDDVILV GGQTRMPMVQ KKVAEFFGKEPRKDVNPDEA VAIGAAVQGG VLTG.
Product Science Overview
DnaK is a well-studied molecular chaperone from the heat shock protein 70 (Hsp70) family, found in Escherichia coli (E. coli). It plays a crucial role in protein folding, assembly, and disassembly of protein complexes, and in the response to stress conditions such as heat shock . The ATPase binding domain of DnaK is particularly important for its function, as it binds and hydrolyzes ATP, driving conformational changes necessary for its chaperone activity .
The DnaK protein consists of three major functional domains:
- N-terminal ATPase Domain: This domain binds ATP and hydrolyzes it to ADP, providing the energy required for the chaperone activity .
- Substrate Binding Domain: This domain binds to unfolded or misfolded protein substrates, preventing their aggregation and assisting in their proper folding.
- C-terminal Domain: This domain is involved in the regulation of the chaperone activity and interaction with co-chaperones.
The ATPase domain of DnaK, specifically, is responsible for the binding and hydrolysis of ATP, which is essential for the chaperone cycle. This domain undergoes conformational changes upon ATP binding and hydrolysis, which are transmitted to the substrate binding domain, facilitating the binding and release of substrate proteins .
Recombinant production of the DnaK ATPase binding domain involves cloning the gene encoding this domain into an E. coli expression vector. The protein is then expressed in E. coli cells and purified using conventional column chromatography techniques . This recombinant protein is used in various biochemical and biophysical studies to understand the mechanism of ATP hydrolysis and its role in the chaperone activity of DnaK.
The recombinant DnaK ATPase binding domain is widely used in research to study:
- Protein Folding: Understanding how molecular chaperones assist in the folding of newly synthesized or stress-denatured proteins.
- Protein-Protein Interactions: Investigating the role of DnaK in the assembly and disassembly of protein complexes.
- Stress Response: Exploring the mechanisms by which cells respond to stress conditions, such as heat shock, and the role of chaperones in this process .
