REN Human, HEK
Description
Product Specs
Product Science Overview
Renin is an enzyme that plays a crucial role in the renin-angiotensin-aldosterone system (RAAS), which regulates blood pressure and fluid balance in the body. It is produced by the juxtaglomerular cells of the kidney and is responsible for the conversion of angiotensinogen to angiotensin I, a precursor to the potent vasoconstrictor angiotensin II. The study and manipulation of renin have significant implications for the treatment of hypertension and related cardiovascular diseases.
Recombinant human renin is produced using genetic engineering techniques, where the gene encoding human renin is inserted into a host cell line, such as Human Embryonic Kidney (HEK) 293 cells. This allows for the production of large quantities of renin for research and therapeutic purposes.
The preparation of recombinant human renin involves several steps:
- Gene Cloning: The cDNA encoding the full length of native human preprorenin (406 amino acid residues) is introduced into the HEK-293 cell line .
- Cell Culture: A clonal cell line expressing prorenin is generated and grown under serum-free conditions in a hollow fiber bioreactor .
- Purification: Prorenin is constitutively secreted and purified directly from the conditioned medium. Concanavalin A chromatography effectively enriches and purifies prorenin to 90% homogeneity in a single step . Prorenin is then converted to active renin by trypsin digestion to remove the propeptide. Active renin is further purified using a cation exchange column followed by a gel filtration column .
Biochemical characterization of the recombinant enzyme shows that both binding and catalytic properties are essentially identical to previously reported activities for purified renin . This high level of purity and activity makes recombinant human renin suitable for supporting structural biology efforts, such as X-ray crystallography studies .
Recombinant human renin is used in various applications, including:
- Enzyme Kinetics: Studying the enzyme kinetics of renin to understand its catalytic mechanism and identify potential inhibitors .
- Drug Screening: Screening for inhibitors that can block the activity of renin, which is a therapeutic target for hypertension .
- Structural Studies: Supporting structural biology efforts to determine the three-dimensional structure of renin, which can aid in the design of novel inhibitors .
