DnaK SBD
Greater than 95.0% as determined by SDS-PAGE.
Description
Recombinant DnaK Substrate Binding domain produced in E.Coli is a single, non-glycosylated polypeptide chain containing (385-546 a.a.) 163 amino acids and having a molecular mass of 17.7 kDa.
Product Specs
MDVKDVLLLD VTPLSLGIET MGGVMTTLIA KNTTIPTKHS QVFSTAEDNQ SAVTIHVLQG ERKRAADNKS LGQFNLDGIN PAPRGMPQIE VTFDIDADGI LHVSAKDKNS GKEQKITIKA SSGLNEDEIQ KMVRDAEANA EADRKFEELV QTRNQGDHLL HST.
Product Science Overview
The DnaK protein, also known as Heat Shock Protein 70 (Hsp70), is a molecular chaperone found in Escherichia coli (E. coli). It plays a crucial role in protein folding, assembly, and preventing the aggregation of stress-denatured proteins. The DnaK Substrate Binding Domain (SBD) is a specific region of the DnaK protein that is essential for its chaperone activity.
The DnaK protein consists of three main domains:
- Nucleotide Binding Domain (NBD): Responsible for ATP binding and hydrolysis.
- Substrate Binding Domain (SBD): Binds to unfolded or partially folded polypeptides.
- C-terminal Domain: Involved in the regulation of the chaperone activity.
The Substrate Binding Domain (SBD) of DnaK, specifically residues 508-638, is α-helical and acts as a lid covering the substrate binding cleft . This domain is critical for recognizing and binding to unfolded or misfolded proteins, thereby preventing their aggregation and assisting in their proper folding.
Recombinant DnaK Substrate Binding Domain is produced in E. coli as a single, non-glycosylated polypeptide chain. The recombinant protein typically contains 163 amino acids and has a molecular mass of approximately 17.7 kDa . The production process involves cloning the gene encoding the DnaK SBD into an expression vector, transforming E. coli cells with this vector, and inducing protein expression. The recombinant protein is then purified using conventional column chromatography techniques .
The recombinant DnaK Substrate Binding Domain has several applications in biochemical and biophysical research:
- Protein Folding Studies: Used to study the mechanisms of protein folding and the role of molecular chaperones.
- Protein-Protein Interactions: Investigating interactions between DnaK and its substrate proteins.
- Drug Discovery: Screening for potential inhibitors or modulators of DnaK activity, which could be useful in developing treatments for diseases related to protein misfolding and aggregation.
The recombinant DnaK SBD is typically formulated in a buffer containing Tris-HCl, β-mercaptoethanol (B-ME), and dithiothreitol (DTT) to maintain its stability . It should be stored at 4°C for short-term use (2-4 weeks) and at -20°C for long-term storage. To prevent degradation, it is recommended to avoid multiple freeze-thaw cycles and to add a carrier protein such as human serum albumin (HSA) or bovine serum albumin (BSA) for long-term storage .
