GroEL E.Coli
Description
Recombinant GroEL produced in E.Coli is a single, non-glycosylated polypeptide chain containing 548 amino acids (1-548) and having a molecular mass of 57.3kDa. GroEL is purified by proprietary chromatographic techniques.
Product Specs
Product Science Overview
GroEL is composed of 14 identical subunits arranged in two stacked heptameric rings, forming a cylindrical structure. Each subunit has a molecular mass of approximately 57 kDa. The central cavity of the GroEL complex provides an isolated environment where unfolded or partially folded polypeptides can achieve their native conformation.
GroEL works in conjunction with its co-chaperonin, GroES (HSP10), which forms a lid-like structure over the GroEL cavity. The binding and hydrolysis of ATP drive conformational changes in GroEL, facilitating the encapsulation and release of substrate proteins. This ATP-dependent mechanism ensures that proteins are folded correctly and efficiently.
Recombinant GroEL (HSP60) from E. coli is produced using genetic engineering techniques. The GroEL gene is amplified by polymerase chain reaction (PCR) and cloned into an expression vector. This vector is then introduced into E. coli cells, which express the GroEL protein. The recombinant protein is subsequently purified using chromatographic techniques to achieve high purity levels .
Recombinant GroEL (HSP60) has several applications in research and biotechnology:
- Protein Folding Studies: GroEL is used to study the mechanisms of protein folding and the role of chaperonins in this process.
- Structural Biology: The well-defined structure of GroEL makes it a model system for studying protein-protein interactions and conformational changes.
- Biotechnology: GroEL can be used to improve the yield and quality of recombinant proteins by assisting in their proper folding during expression in E. coli.
- Medical Research: Understanding the function of GroEL and its human homologs can provide insights into diseases related to protein misfolding and aggregation, such as neurodegenerative disorders.
Recombinant GroEL (HSP60) is typically stored in a buffer containing Tris-HCl, NaCl, DTT, and glycerol to maintain its stability. It should be stored at 4°C for short-term use 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 .
