DnaK Lid Covering Substrate E.Coli
(b) Analysis by SDS-PAGE.
Description
Product Specs
MNEDEIQKMV RDAEANAEAD RKFEELVQTR NQGDHLLHST RKQVEEAGDK LPADDKTAIESALTALETAL KGEDKAAIEA KMQELAQVSQ KLMEIAQQQH AQQQTAGADASANNAKDDDVVDAEFEEVKDKK.
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 the folding and assembly of newly synthesized polypeptide chains and in preventing the aggregation of stress-denatured proteins. The DnaK protein is essential for the survival of bacteria under stress conditions, such as high temperatures.
The DnaK protein consists of several domains, including the substrate-binding domain (SBD) and the nucleotide-binding domain (NBD). The SBD is responsible for binding to unfolded or partially folded polypeptides, while the NBD binds and hydrolyzes ATP, providing the energy required for the chaperone activity.
The DnaK Lid Covering Substrate domain, specifically, refers to a segment of the DnaK protein that acts as a lid covering the substrate-binding cleft. This lid is composed of residues 508-638 and is α-helical in structure . The lid plays a critical role in regulating the binding and release of substrate proteins.
Recombinant DnaK Lid Covering Substrate domain is produced in E. coli as a single, non-glycosylated polypeptide chain containing 132 amino acids and having a molecular mass of 14.6 kDa . The recombinant protein is purified to apparent homogeneity using conventional column chromatography techniques . The protein is typically formulated in a buffer containing 25mM Tris-HCl (pH 7.5), 100mM NaCl, 5mM DTT, and 10% glycerol .
The recombinant DnaK Lid Covering Substrate domain is used in various research applications, including:
- Protein Folding Studies: Understanding the mechanisms of protein folding and the role of molecular chaperones.
- Stress Response Research: Investigating how bacteria respond to stress conditions and the role of heat shock proteins in stress tolerance.
- Drug Development: Screening for potential inhibitors of DnaK activity, which could serve as antibacterial agents.
The recombinant DnaK Lid Covering Substrate domain should be stored at 4°C if used within 2-4 weeks. For longer storage periods, it should be frozen at -20°C, and it is recommended to add a carrier protein (0.1% HSA or BSA) to prevent degradation . Multiple freeze-thaw cycles should be avoided to maintain protein integrity.
