CASP3 Human, Sf9
CASP3, CPP32, CPP32B, SCA-1, CASP-3, Apopain, Cysteine protease CPP32, CPP-32, Protein Yama, SREBP cleavage activity 1.
Description
CASP3 Human produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 256 amino acids (29-277 a.a.) and having a molecular mass of 29.4kDa (Migrates at 13.5-18kDa on SDS-PAGE under reducing conditions).
CASP3 is expressed with a 6 amino acid His tag at C-Terminus and purified by proprietary chromatographic techniques.
Product Specs
Caspase 3, also known as Apoptosis-Related Cysteine Peptidase (CASP3), is a protein belonging to the cysteine-aspartic acid protease (caspase) family. These proteases play a crucial role in the execution phase of programmed cell death, or apoptosis. Caspases are initially present in cells as inactive precursors called proenzymes. Upon receiving specific signals, these proenzymes undergo a series of cleavages at conserved aspartic acid residues. This processing generates two subunits, a large and a small subunit, which then combine to form the active caspase enzyme. CASP3, once activated, can further activate other caspases, including caspases 6, 7, and 9, amplifying the apoptotic cascade. Conversely, CASP3 itself can be activated by caspases 8, 9, and 10. This intricate activation network highlights the tightly regulated nature of apoptosis. CASP3 has been implicated in several key cellular processes, including: 1. Cleavage of Amyloid-beta 4A Precursor Protein: CASP3 is a major enzyme responsible for cleaving the amyloid-beta 4A precursor protein, a process linked to neuronal death in Alzheimer's disease. 2. Cleavage of Huntingtin: CASP3 is involved in the breakdown of huntingtin, a protein whose mutation is associated with Huntington's disease. 3. Cleavage of Sterol Regulatory Element Binding Proteins (SREBPs): CASP3 can cleave SREBPs, transcription factors involved in lipid metabolism, separating their DNA-binding domain from the membrane attachment domain. 4. Initiation of Cell Adhesion: In sympathetic neurons, CASP3 plays a role in initiating cell adhesion by cleaving the RET receptor tyrosine kinase.
This product consists of the human CASP3 protein produced using Sf9 insect cells infected with a baculovirus expression system. It is a single polypeptide chain that has undergone glycosylation, a common post-translational modification. The protein encompasses amino acids 29 to 277 of the CASP3 sequence, resulting in a protein of 256 amino acids. The molecular weight of the protein is 29.4 kDa. On SDS-PAGE analysis under reducing conditions, it migrates with an apparent molecular weight between 13.5 and 18 kDa. The protein has been engineered to include a 6-amino acid histidine tag at the C-terminus, facilitating its purification. Purification is achieved using proprietary chromatographic techniques.
The CASP3 protein is provided in a solution at a concentration of 0.5 mg/ml. The solution is buffered with 20 mM HEPES at pH 7.5 and contains 0.1 M NaCl, 1 mM EDTA, 20% glycerol, and 1 mM DTT.
For short-term storage (up to four weeks), the CASP3 protein solution can be stored at 4°C. For extended storage, it is recommended to store the protein at -20°C. To ensure optimal stability during long-term storage, consider adding a carrier protein such as albumin (HSA or BSA) to a final concentration of 0.1%. It is crucial to minimize freeze-thaw cycles to maintain protein integrity.
The purity of the CASP3 protein is greater than 90%, as determined by SDS-PAGE analysis.
The specific activity of the CASP3 protein is greater than 5,000 pmol/min/µg. This activity is determined by measuring the enzyme's ability to cleave a specific substrate, Ac-DEVD-AFC, into Ac-DEVD and AFC. One unit of enzyme activity is defined as the amount of enzyme required to release 1 pmole of AFC per minute at a pH of 7.5 and a temperature of 25°C.
CASP3, CPP32, CPP32B, SCA-1, CASP-3, Apopain, Cysteine protease CPP32, CPP-32, Protein Yama, SREBP cleavage activity 1.
MSGISLDNSY KMDYPEMGLC IIINNKNFHK STGMTSRSGT DVDAANLRET FRNLKYEVRN
KNDLTREEIV ELMRDVSKED HSKRSSFVCV LLSHGEEGII FGTNGPVDLK KITNFFRGDR
CRSLTGKPKL FIIQACRGTE LDCGIETDSG VDDDMACHKI PVEADFLYAY STAPGYYSWR
NSKDGSWFIQ SLCAMLKQYA DKLEFMHILT RVNRKVATEF ESFSFDATFH AKKQIPCIVS MLTKELYFYH HHHHHH
Product Science Overview
Caspases are classified into two main categories based on their roles: initiator caspases (e.g., caspase-8 and caspase-9) and executioner caspases (e.g., caspase-3, caspase-6, and caspase-7) . Caspase 3 is an executioner caspase, meaning it is responsible for the cleavage of various key cellular proteins, leading to the morphological and biochemical changes observed during apoptosis .
Caspase 3 exists as an inactive proenzyme (procaspase) that undergoes proteolytic processing at conserved aspartic residues to produce two subunits, large and small, which dimerize to form the active enzyme . The active site of caspase 3 contains a cysteine residue (Cys-163) and a histidine residue (His-121), which are crucial for its enzymatic activity .
Caspase 3 is involved in the cleavage and activation of other caspases, such as caspase-6 and caspase-7, and is itself activated by caspases-8, -9, and -10 . It plays a central role in the execution phase of apoptosis by cleaving various cellular substrates, including poly (ADP-ribose) polymerase (PARP), which is involved in DNA repair, and the amyloid-beta precursor protein, which is associated with neuronal death in Alzheimer’s disease .
The activation of caspase 3 can occur through two main apoptotic pathways: the intrinsic (mitochondrial) pathway and the extrinsic (death receptor) pathway . In the intrinsic pathway, cytochrome c released from mitochondria binds to Apaf-1 and procaspase-9 to form the apoptosome, leading to the activation of caspase-9, which in turn activates caspase 3 . In the extrinsic pathway, the binding of ligands to death receptors (e.g., Fas) leads to the formation of the death-inducing signaling complex (DISC), which activates caspase-8, subsequently activating caspase 3 .
Caspase 3 activity is tightly regulated by various mechanisms to ensure controlled apoptosis. Inhibitors of apoptosis proteins (IAPs) can bind to and inhibit active caspases, including caspase 3 . Additionally, the expression of caspase 3 can be regulated at the transcriptional level by various factors, including p53, a tumor suppressor protein that can induce apoptosis in response to DNA damage .
Human recombinant caspase 3 produced in Sf9 insect cells is used for research purposes to study its structure, function, and role in apoptosis. The recombinant enzyme retains the same biological properties and functions as the native enzyme, making it a valuable tool for investigating the mechanisms of apoptosis and for developing potential therapeutic interventions targeting caspase 3 .
