METAP2/Methionine aminopeptidase 2 蛋白, Human (sf9, His)

METAP2/Methionine aminopeptidase 2 protein removes N-terminal methionine from nascent proteins. It preferentially cleaves when the second residue is small and uncharged. METAP2 has higher catalytic activity for Met-Val peptides than METAP1, suggesting its role in processing Met-Val and Met-Thr sequences in vivo. It also protects EIF2S1 from phosphorylation by inhibitory kinases, regulating protein synthesis. METAP2/Methionine aminopeptidase 2 Protein, Human (sf9, His) is the recombinant human-derived METAP2/Methionine aminopeptidase 2 protein, expressed by sf9 insect cells , with N-6*His labeled tag.

METAP2/甲硫氨酸氨基肽酶 2 蛋白通过共翻译从新生蛋白中去除 N 末端甲硫氨酸来发挥作用。当一级序列中的第二个残基较小且不带电荷（例如 Met-Ala-、Cys、Gly、Pro、Ser、Thr 或 Val）时，就会发生这种 N 端甲硫氨酸裂解。人 METAP2 对 Met-Val 肽的催化活性显着高于 METAP1，表明其负责体内加工含有 N 端 Met-Val 和 Met-Thr 序列的蛋白质。此外，METAP2 还可以保护真核起始因子 EIF2S1 免受 EIF2AK2/PKR 和 EIF2AK1/HCR 等抑制性激酶的翻译抑制性磷酸化，从而在调节蛋白质合成中发挥关键作用。

Measure by its ability to remove methionine from a fluorogenic peptide substrate H-Met-Gly-Pro-AMC. The resulting GP-AMC is cleaved by Recombinant Human DPPIV/CD26 that incubate at room temperature in kinetic mode for 5 minutes. The specific activity is >15 pmol/min/µg.

Materials
Assay buffer: 50 mM HEPES, 0.1 mM CoCl₂, 100 mM NaCl, pH 7.5
METAP2/Methionine aminopeptidase 2 Protein, Human (sf9, His) (HY-P70364)
CD26/Dipeptidyl Peptidase 4 Protein, Human (HEK293, His) (HY-P70017)
Substrate: Met-Gly-Pro-AMC (HY-P4344)
Standard: 7-Amino-4-Methyl-Courmarin (HY-D0027)

Procedure
1. Standard curve: Dilute standard to 0, 6.25, 12.5, 25, 50, 100 μmol/L with assay buffer, and add 100 μL of each into the black zymography well. The excitation and emission wavelengths were 380 nm and 460 nm, respectively, and read in kinetic mode for 5 min. The measured RFU was used as the horizontal coordinate, and the amount of standard substance was used as the vertical coordinate to make a standard curve and obtain the standard curve formula.
2. Dilute Human METAP2 to 50 μg/mL with assay buffer.
3. Dilute the substrate to 500 μM with 2 μg/mL Human CD26 in assay buffer.
4. Experimental group: 50 μg/mL of Human METAP2L 50 μL was loaded into the plate and 50 μL of substrate/Human CD26 mixture was added to start the reaction.
Control group: 50 μL substrate/Human CD26 mixture + 50 μL buffer.
5. Incubate for 10 minutes at room temperature.
6. Read in kinetic mode for 5 minutes at excitation and emission wavelengths of 380 nm and 460 nm, respectively.
7. Calculate the specific activity:

Human

Sf9 insect cells

N-6*His

P50579-1 (A2-Y478)

10988  [NCBI]

Full Length of Isoform-1 Mature Protein

AGVEEVAASGSHLNGDLDPDDREEGAASTAEEAAKKKRRKKKKSKGPSAAGEQEPDKESGASVDEVARQLERSALEDKERDEDDEDGDGDGDGATGKKKKKKKKKRGPKVQTDPPSVPICDLYPNGVFPKGQECEYPPTQDGRTAAWRTTSEEKKALDQASEEIWNDFREAAEAHRQVRKYVMSWIKPGMTMIEICEKLEDCSRKLIKENGLNAGLAFPTGCSLNNCAAHYTPNAGDTTVLQYDDICKIDFGTHISGRIIDCAFTVTFNPKYDTLLKAVKDATNTGIKCAGIDVRLCDVGEAIQEVMESYEVEIDGKTYQVKPIRNLNGHSIGQYRIHAGKTVPIVKGGEATRMEEGEVYAIETFGSTGKGVVHDDMECSHYMKNFDVGHVPIRLPRTKHLLNVINENFGTLAFCRRWLDRLGESKYLMALKNLCDLGIVDPYPPLCDIKGSYTAQFEHTILLRPTCKEVVSRGDDY

Approximately 58-70 kDa due to the glycosylation.

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

溶液.

Supplied as a 0.22 μm filtered solution of 20 mM Tris, 500 mM NaCl, 10% glycerol, pH 8.0.
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.