THOP1 Protein, Human (His)
Based on 1 Customer Validation
The THOP1 protein metabolizes short neuropeptides, degrading them in the cytoplasm. It also degrades amyloid precursor protein, producing fragments associated with amyloid production. THOP1 Protein, Human (His) is the recombinant human-derived THOP1 protein, expressed by E. coli , with C-6*His labeled tag.
- Species: Human
- Source: E. coli
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Storage:Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.
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Biological Activity
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Technical Parameters
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Product Properties
- Documentation
- Help & FAQs
Biological Activity
The THOP1 protein metabolizes short neuropeptides, degrading them in the cytoplasm. It also degrades amyloid precursor protein, producing fragments associated with amyloid production. THOP1 Protein, Human (His) is the recombinant human-derived THOP1 protein, expressed by E. coli , with C-6*His labeled tag.
THOP1 Protein plays a crucial role in the metabolism of neuropeptides with a length of fewer than 20 amino acid residues, participating specifically in cytoplasmic peptide degradation. Its enzymatic activity extends to the degradation of the amyloid-beta precursor protein, generating fragments associated with amyloidogenesis. Additionally, THOP1 serves as a regulatory component in the cannabinoid signaling pathway, where it facilitates the degradation of hemopressin, an antagonist peptide targeting the cannabinoid receptor CNR1. The diverse functions of THOP1 underscore its involvement in intricate cellular processes, ranging from neuropeptide metabolism to the modulation of key signaling pathways.
Measured by its ability to cleave a fluorogenic peptide substrate Mca-PLGPK(Dnp)-OH that incubate at room temperature in kinetic mode for 5 minutes. The specific activity is 2815.75 pmol/min/µg.
Assay Procedure
Materials
Assay buffer: 25 mM Tris, 150 mM NaCl, pH 7.5
THOP1 Protein, Human (His) (HY-P71359)
Substrate: Mca-Pro-Leu-Gly-Pro-D-Lys(Dnp) (HY-P3792), dissolved in DMSO to 2 mM
Standard: MCA-Pro-Leu-OH
Procedure
1. Preparation of standard curve: Dilute MCA-Pro-Leu-OH to 100, 50, 25, 12.5, 6.25, 3.125, 1.5625, and 0 μmol/L respectively with assay buffer. Add 100 μL of each concentration standard to a 96-well plate. Read at wavelengths of 320 nm and 405 nm in kinetic mode (top read) for 5 minutes. Plot a standard curve with the standard concentration as the abscissa and the measured OD as the ordinate, and obtain the standard curve equation.
2. Dilute Human THOP1 to 500 ng/mL with assay buffer.
5. Dilute the substrate to 20 μM with assay buffer.
6. Add 50 μL of Human THOP1 to a 96-well plate, and add 50 μL of 20 μM substrate to initiate the reaction. For blank wells, add 50 μL of assay buffer and 50 μL of 20 μM substrate.
7. Read at wavelengths of 320 nm and 405 nm in kinetic mode (top read) for 5 minutes, respectively.
8. Calculate the specific activity:
Specific Activity (pmol/min/μg) = | Adjusted Vmax* (RFU/min) x Conversion Factor ** (pmol/RFU) |
| amount of enzyme (μg) |
*Adjusted for Substrate Blank
**Derived using calibration standard
Per Well:
Human THOP1: 0.025 μg
Substrate: 10 μM
Technical Parameters
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Species Human
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Source E. coli
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Tag C-6*His
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Accession
P52888 (K2-C689)
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Molecular Construction
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N-term
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THOP1 (K2-C689)
Accession # P52888 -
6*His
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C-term
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Protein Length
Partial
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Synonyms
THOP1; MP78; Thimet Oligopeptidase 1; MEPD_HUMAN; Thimet Oligopeptidase; EP24.15; Endopeptidase 24.15; TOP
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AA Sequence
KPPAACAGDMADAASPCSVVNDLRWDLSAQQIEERTRELIEQTKRVYDQVGTQEFEDVSYESTLKALADVEVTYTVQRNILDFPQHVSPSKDIRTASTEADKKLSEFDVEMSMREDVYQRIVWLQEKVQKDSLRPEAARYLERLIKLGRRNGLHLPRETQENIKRIKKKLSLLCIDFNKNLNEDTTFLPFTLQELGGLPEDFLNSLEKMEDGKLKVTLKYPHYFPLLKKCHVPETRRKVEEAFNCRCKEENCAILKELVTLRAQKSRLLGFHTHADYVLEMNMAKTSQTVATFLDELAQKLKPLGEQERAVILELKRAECERRGLPFDGRIRAWDMRYYMNQVEETRYCVDQNLLKEYFPVQVVTHGLLGIYQELLGLAFHHEEGASAWHEDVRLYTARDAASGEVVGKFYLDLYPREGKYGHAACFGLQPGCLRQDGSRQIAIAAMVANFTKPTADAPSLLQHDEVETYFHEFGHVMHQLCSQAEFAMFSGTHVERDFVEAPSQMLENWVWEQEPLLRMSRHYRTGSAVPRELLEKLIESRQANTGLFNLRQIVLAKVDQALHTQTDADPAEEYARLCQEILGVPATPGTNMPATFGHLAGGYDAQYYGYLWSEVYSMDMFHTRFKQEGVLNSKVGMDYRSCILRPGGSEDASAMLRRFLGRDPKQDAFLLSKGLQVGGCEPEPQVC
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Predicted Molecular Mass
80 kDa
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Molecular Weight
Approximately 85 kDa, based on SDS-PAGE under reducing conditions.
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Purity
≥ 95%, as determined by reducing SDS-PAGE.
Product Properties
Solution
Supplied as a 0.22 μm filtered solution of 20 mM Tris-HCl, 500 mM NaCl, 50% Glycerol, pH 7.4.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.
<1 EU/μg, determined by LAL method.
Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.
Shipping with dry ice.
Documentation
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Data Sheet (239 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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