SOD1 recombinant proteins refer to engineered versions of Superoxide Dismutase 1 (SOD1), an important antioxidant enzyme that protects cells from oxidative damage by catalyzing the dismutation of superoxide radicals into oxygen and hydrogen peroxide. Mutations in the SOD1 gene are linked to Amyotrophic Lateral Sclerosis (ALS), making SOD1 a critical target for research into neurodegenerative diseases and oxidative stress. Recombinant SOD1 proteins are used for studying enzyme function, disease mechanisms, and potential therapeutic approaches.

Structure and Function

• Superoxide Dismutase 1 (SOD1) is a 16 kDa enzyme composed of approximately 154 amino acids. It contains a crucial metal-binding site, typically copper and zinc, which is essential for its enzymatic activity.
• SOD1 plays a key role in protecting cells from oxidative stress by converting superoxide radicals (O₂⁻) into less reactive molecules, oxygen (O₂) and hydrogen peroxide (H₂O₂).
• Recombinant SOD1 proteins often include the full-length enzyme or specific mutants to study structural and functional aspects, including those associated with ALS.

Production

• Recombinant SOD1 proteins are produced in expression systems such as E. coli, yeast, insect cells, or mammalian cells. The choice of system depends on the need for proper protein folding and metal incorporation.
• The gene encoding SOD1 is cloned into an expression vector, and the protein is expressed and purified using affinity chromatography. Common tags like His-tag or GST-tag are used to facilitate purification.
• For proper enzymatic activity, metal cofactors (copper and zinc) may be added during the purification process or during in vitro refolding.

Applications

• Basic Research: Recombinant SOD1 proteins are used to study the enzyme's role in oxidative stress, its mechanism of action, and the impact of genetic mutations. This research helps in understanding how SOD1 mutations contribute to ALS and other neurodegenerative diseases.
• Disease Modeling: SOD1 proteins, including various mutants linked to ALS, are used in experimental models to investigate disease mechanisms and to test potential therapies.
• Therapeutic Development: Understanding the structure-function relationship of SOD1 and the effects of mutations can aid in developing drugs or gene therapies aimed at treating or mitigating ALS and other conditions related to oxidative stress.
• Diagnostics: Recombinant SOD1 proteins are used to develop diagnostic assays for detecting SOD1 mutations or assessing oxidative stress levels in biological samples.

Validation and Quality Control

• Validation involves confirming the identity, purity, and activity of recombinant SOD1 proteins through techniques such as SDS-PAGE, Western blotting, and mass spectrometry.
• Functional assays, including enzyme activity assays that measure superoxide dismutation, are performed to ensure that the recombinant proteins exhibit the expected enzymatic activity and that any mutations are properly characterized