Recombinant Human Methionine Aminopeptidase 2 Protein, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human Methionine Aminopeptidase 2 Protein, CF Summary

• Details of Functionality: Measure by its ability to remove methionine from a fluorogenic peptide substrate H-Met-Gly-Pro-AMC (Catalog # ES017). The resulting GP-AMC is cleaved by Recombinant Human DPPIV/CD26 (Catalog # 9168-SE). The specific activity is >15 pmol/min/µg, as measured using the described conditions.
• Source: Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Methionine Aminopeptidase 2/METAP2 protein
  Ala2-Tyr478, with a C-terminal 10-His tag
• Accession #: P50579
• N-terminal Sequence: Ala2
• Protein/Peptide Type: Recombinant Enzymes
• Gene: METAP2
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 54 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 66 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Assay Procedure:
  • Assay Buffer: 50 mM HEPES, 0.1 mM CoCl₂, 100 mM NaCl, pH 7.5
  • Recombinant Human Methionine Aminopeptidase 2/METAP2 (rhMETAP2) (rhMETAP2) (Catalog # 3795-ZN
  • Recombinant Human DPPIV/CD26 (rhCD26) (Catalog # 9168-SE)
  • Substrate: Met-Gly-Pro-AMC (Catalog # ES017)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhMETAP2 to 10 µg/mL in Assay Buffer.
  2. Dilute Substrate to 500 µM with 2 µg/mL rhCD26 in Assay Buffer.
  3. Load 50 µL of 10 µg/mL rhMETAP2 into a plate, and start the reaction by adding 50 µL of Substrate/rhCD26 mixture.
  4. Incubate at room temperature for 10 minutes.
  5. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes (effectively reading timepoints between 10 and 15 minutes).
  6. Calculate specific activity using the following formula:

  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 7-Amino-4-Methyl-Courmarin (AMC) (Sigma, Catalog#: A-9891).

  Per Well:
  • rhMETAP2: 0.500 µg
  • rhCD26: 0.100 µg
  • Substrate: 250 µM

Notes

This product is covered by US Patent Nos. 5,885,820 and 5,888,796 licensed from Mediomics, LLC.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Methionine Aminopeptidase 2 Protein, CF

• Amp2
• eIF-2-associated p67 homolog
• Initiation factor 2-associated 67 kDa glycoprotein
• MAP 2
• MAP2
• MetAP 2
• METAP2
• Methionine Aminopeptidase 2
• methionyl aminopeptidase 2
• MNPEP
• p67
• p67EC 3.4.11.18
• p67eIF2
• Peptidase M 2

Background

The human METAP2 gene encodes Methionine Aminopeptidase 2, a member of the M24 family of metalloproteases. METAPs catalyze the removal of the initiator methionine residue from nascent peptides (1) and are essential for cell growth (2). METAP2 plays an important role in the development of different types of cancer (3) and has been a novel target for developing anti-cancer drugs.

1. Lowther, W.T. and B.W. Matthews (2000) Biochim. Biophys. Acta 1477:157.
2. Li, X. and Y.H. Chang (1995) Proc. Natl. Acad. Sci. USA 92:12357.
3. Selvakumar, P. et al. (2006) Biochim. Biophys. Acta 1765:148.

Publications for Methionine Aminopeptidase 2/METAP2 (3795-ZN)(3)

Publications using 3795-ZN

• ZS Clauss, CL Wardzala, AE Schlirf, NS Wright, SS Saini, B Onoa, C Bustamante, JR Kramer Tunable, biodegradable grafting-from glycopolypeptide bottlebrush polymers Nature Communications, 2021-11-09;12(1):6472. 2021-11-09 [PMID: 34753949] (Bioassay, Human)
• E W?glarz-To, M Talma, M Giurg, HV Westerhoff, R Janowski, A Mucha Neutral metalloaminopeptidases APN and MetAP2 as newly discovered anticancer molecular targets of actinomycin D and its simple analogs Oncotarget, 2018-06-29;9(50):29365-29378. 2018-06-29 [PMID: 30034623] (Enzyme Assay, Human)
• Chiu J, Wong J, Hogg P Redox regulation of methionine aminopeptidase 2 activity. J Biol Chem, 2014-04-03;289(21):15035-43. 2014-04-03 [PMID: 24700462] (Bioassay)

Bioinformatics

• Gene Symbol: METAP2
• Uniprot:
  • P50579()