THOP1 蛋白, Human (His)

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蛋白在长度小于20个氨基酸残基的神经肽的代谢中发挥着至关重要的作用，特别参与细胞质肽的降解。其酶活性延伸至淀粉样β前体蛋白的降解，产生与淀粉样蛋白生成相关的片段。此外，THOP1 作为大麻素信号通路中的调节成分，促进加压素（一种针对大麻素受体 CNR1 的拮抗剂肽）的降解。THOP1 的多种功能强调了它参与复杂的细胞过程，从神经肽代谢到关键信号通路的调节。

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.

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:

Human

E. coli

C-6*His

P52888 (K2-C689)

7064  [NCBI]

Partial

KPPAACAGDMADAASPCSVVNDLRWDLSAQQIEERTRELIEQTKRVYDQVGTQEFEDVSYESTLKALADVEVTYTVQRNILDFPQHVSPSKDIRTASTEADKKLSEFDVEMSMREDVYQRIVWLQEKVQKDSLRPEAARYLERLIKLGRRNGLHLPRETQENIKRIKKKLSLLCIDFNKNLNEDTTFLPFTLQELGGLPEDFLNSLEKMEDGKLKVTLKYPHYFPLLKKCHVPETRRKVEEAFNCRCKEENCAILKELVTLRAQKSRLLGFHTHADYVLEMNMAKTSQTVATFLDELAQKLKPLGEQERAVILELKRAECERRGLPFDGRIRAWDMRYYMNQVEETRYCVDQNLLKEYFPVQVVTHGLLGIYQELLGLAFHHEEGASAWHEDVRLYTARDAASGEVVGKFYLDLYPREGKYGHAACFGLQPGCLRQDGSRQIAIAAMVANFTKPTADAPSLLQHDEVETYFHEFGHVMHQLCSQAEFAMFSGTHVERDFVEAPSQMLENWVWEQEPLLRMSRHYRTGSAVPRELLEKLIESRQANTGLFNLRQIVLAKVDQALHTQTDADPAEEYARLCQEILGVPATPGTNMPATFGHLAGGYDAQYYGYLWSEVYSMDMFHTRFKQEGVLNSKVGMDYRSCILRPGGSEDASAMLRRFLGRDPKQDAFLLSKGLQVGGCEPEPQVC

80-85 kDa

Greater than 95% as determined by reducing SDS-PAGE.

溶液.

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.

N/A.

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.