Recombinant Human CAT protein(Ala2-Leu527), His-tagged

• Cat.No.: CAT-2500H
• Product Overview: Recombinant Human CAT (P04040) (Ala 2-Leu 527) was expressed in Insect Cells, with a polyhistidine tag at the N-terminus.

Specification

• Species: Human
• Source: Insect Cells
• Tag: His
• Protein Length: 2-527 a.a.
• Form: Lyophilized from sterile 50mM Tris, 100mM NaCl, pH 8.0, 10% gly. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween80 are added as protectants before lyophilization.
• Molecular Mass: The recombinant human CAT consists of 545 amino acids and predicts a molecular mass of 61.9 kDa. It migrates as an approximately 60 kDa band in SDS-PAGE under reducing conditions.
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method
• Purity: > 80 % as determined by SDS-PAGE
• Storage: Samples are stable for up to twelve months from date of receipt at -20°C to -80°C. Store it under sterile conditions at -20°C to -80°C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.

Gene Information

• Gene Name: CAT catalase [ Homo sapiens ]
• Official Symbol: CAT
• Synonyms: CAT; catalase; MGC138422; MGC138424
• Gene ID: 847
• mRNA Refseq: NM_001752
• Protein Refseq: NP_001743
• MIM: 115500
• UniProt ID: P04040

Case Study

Case 1: González-Sánchez MI, et al. Arch Biochem Biophys. 2011

Hydrogen peroxide triggers a redox cycle between methemoglobin and ferrylhemoglobin, causing protein inactivation and oxygen production. This study analyzed how methemoglobin behaves like catalase with H2O2, revealing unusual kinetics tied to protein concentration. Superoxide scavengers increased activity slightly, suggesting mostly biocatalytic oxygen production. The proposed mechanism involves competing reactions affecting hemoglobin radicals, and simulations confirmed that methemoglobin’s catalase-like activity primarily protects it from H2O2-induced inactivation.

Fig1. The effects of the different superoxide scavengers on the catalase-like oxygen production catalyzed by metHb.

Fig2. Time course of O2 production in a catalase-like reaction of metHb (7.5 μM), oxyHb (5.3 μM) and metMb (9.9 μM) in the presence of [H2O2]0 = 2 mM.

Case 2: Lee JN, et al. Biochem Biophys Res Commun. 2018

Peroxisomes play a vital role in cellular metabolism and are controlled by pexophagy, a specific type of autophagy. Catalase, an important enzyme in peroxisomes, breaks down reactive oxygen species (ROS). However, its involvement in pexophagy had not been explored before. This study found that inhibiting catalase, either genetically or with 3-aminotriazole during nutrient deprivation, reduced peroxisomes and key protein levels, increased ROS, and induced NBR1-dependent autophagy. The antioxidant N-acetyl-l-cysteine stopped ROS accumulation and pexophagy, revealing catalase's significant role in managing peroxisomes under nutrient stress.

Fig1. Whole cell lysate from HepG2 cells was analyzed for catalase expression by immunoblotting.

Fig2. HepG2 cells were treated as indicated, immunostained with PMP70 (red), NBR1 (green) and DAPI (blue), and observed under a confocal microscopy.

Application

Recombinant catalase (CAT), an enzyme crafted through genetic engineering, is adept at decomposing hydrogen peroxide into harmless water and oxygen. In research, CAT is vital for shielding cells against oxidative stress, facilitating investigations into cell aging, oxidative damage, and related illnesses. It's also instrumental in understanding genetic factors in conditions sensitive to oxidative stress, such as heart disease and liver cancer. Moreover, in enzyme studies and structural biology, recombinant CAT is key for deciphering its operational mechanisms, with tools like X-ray crystallography enhancing our insight into its function. These insights aid in developing more efficient enzyme formulations. In synthetic biology, CAT is instrumental in creating metabolic pathways and biosensors for monitoring cellular redox states. In industrial applications, recombinant CAT finds use in the food sector for removing hydrogen peroxide residues in milk preservation and cheese production, reducing the need for heat treatment that can diminish nutritional value. In baking, CAT acts as a leavening agent by releasing oxygen. In textiles and paper industries, recombinant CAT helps eliminate hydrogen peroxide in bleaching processes, replacing traditional methods that require high temperatures, thus saving resources and reducing environmental impact. Environmentally, CAT is used to degrade hydrogen peroxide in industrial wastewater, preventing pollution. In pharmaceuticals, recombinant human CAT protein holds potential for developing antioxidant drugs or as an adjunct in cell therapy. Overall, recombinant CAT offers significant potential across research and industrial sectors due to its high efficiency, safety, and eco-friendly nature, making it an essential tool for both scientific study and practical applications.

Fig1. Nanoceria exerts antioxidant effects by scavenging free radicals by mimicking the activities of SOD and CAT. (Da-Long Dong, 2024)