Recombinant Human Transglutaminase 2/TGM2 Protein, CF

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

Recombinant Human Transglutaminase 2/TGM2 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a synthetic peptide Benzyloxycarbonyl-Gln-Gly and NH 2 OH. The specific activity is >1000 pmol/min/ug, as measured under the described condition.
• Source: Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Transglutaminase 2/TGM2 protein Ala2-Ala687 (Asn533Thr and Leu655Val), with an N-terminal Met and 6-His tag
• Accession #: P21980
• N-terminal Sequence: No results obtained
• Protein/Peptide Type: Recombinant Enzymes
• Gene: TGM2
• Purity: >95%, 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: 78 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: 82 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 degreesC as supplied. 3 months, -70 degreesC under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol and DTT.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • Assay Diluent: deionized water
  • Recombinant Human Transglutaminase 2/TGM2 (rhTGM2) (Catalog # 4376-TG)
  • Substrate: Z-Gln-Gly (Sigma, Catalog # C6154), dissolve 500 mM in deionized water, then neutralize it to pH 9.0 with NaOH
  • MES, pH 6.0, 1 M stock
  • DTT (Sigma, Catalog # D-0632), 1 M stock in deionized water
  • CaCl₂ ,1 M stock in deionized water
  • 1 M Hydroxylamine Hydrochloride (Sigma, Catalog # 159417), dissolve in deionized H₂O, then neutralize it to pH 6.0 with NaOH
  • Stop Solution: 0.37 M FeCl₃ (Sigma, Catalog # 236489), 0.67 M HCl, 0.2 M Trichloroacetic Acid
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: Spectramax Plus by Molecular Devices) or equivalent
  1. Prepare Substrate Mixture fresh before running assay. Mix the following components (Dilute all components to the correct concentration in Assay Diluent.):
  1. 10 µL of 500 mM Substrate
  2. 50 µL of 400 mM MES, pH 6.0
  3. 5 µL of 200 mM DTT
  4. 5 µL of 200 mM CaCl₂
  5. 10 µL of 1 M Hydroxylamine Hydrochloride
     (Note: Multiply the volume for each component by the number of reaction vials + 1 to make enough substrate mixture for the assays. For example: If there are 2 reaction vials (including blank), multiply all volumes by 3.)
  2. Dilute rhTGM2 to 0.1 mg/mL in Assay Diluent.
  3. Mix 20 µL of the diluted rhTGM2 with 80 µL Substrate Mixture. For the Substrate Blank mix 20 µL of Assay Diluent with 80 µL Substrate Mixture.
  4. Incubate at 37 °C for 2 hours.
  5. After incubation, stop the reaction with 400 µL of the Stop Solution. Mix well.
  6. Centrifuge at top speed for 2 minutes in a microcentrifuge.
  7. Load 200 µL of the supernatant into a plate.
  8. Read at 525 nm (absorbance) in endpoint mode.
  9. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Abs* (OD) x Conversion Factor** (pmol/OD)) / (Incubation time (min) x amount of enzyme (µg))

  *Adjusted for Substrate Blank
  **Derived using calibration standard L-glutamic acid gamma -monohydroxamate (Sigma, Catalog # G2253).

  Per Well:
  • rhTGM2: 0.8 µg
  • Substrate: 10 mM

Notes

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

Alternate Names for Recombinant Human Transglutaminase 2/TGM2 Protein, CF

• C polypeptide
• EC 2.3.2.13
• G-ALPHA-h
• protein-glutamine gamma-glutamyltransferase 2
• protein-glutamine-gamma-glutamyltransferase
• TG(C)
• TG2
• TGase C
• TGase H
• TGase-2
• TGase-H
• TGC
• TGCGNAH
• TGM2
• Tissue transglutaminase
• transglutaminase 2 (C polypeptide, protein-glutamine-gamma-glutamyltransferase)
• Transglutaminase 2
• Transglutaminase C
• Transglutaminase H
• transglutaminase-2
• tTG

Background

Transglutaminase 2 (TG2), encoded by the TGM2 gene, is also known as tissue transglutaminase (tTG), transgluytaminase C (TGC), and protein-glutamine-gamma -glutamyltransferase. It belongs to the family of transglutaminases that catalyze the posttranslational modification of proteins via calcium dependent cross-linking reactions (1-3). In addition to its function in protein cross-linking, TGM2 is also capable of hydrolyzing both GTP and ATP (4) and has intrinsic kinase activity (5). TGM2 has been implicated in a variety of human diseases including celiac disease, inclusion body myositis, atherosclerosis, and neurodegenerative diseases (6, 7).

1. Gentile, V. et al. (1991) J. Biol. Chem. 266:478.
2. Chen, J.S.K. and K. Mehta (1999) Internat. J. Biochem. Cell Biol. 31:817.
3. Griffin, M. et al. (2002) Biochem. J. 368:377.
4. Lai, T.S. et al. (1998) J. Biol. Chem. 273:1776.
5. Mishra, S. et al. (2007) J. Biol. Chem. 282:18108.
6. Kim, S-Y. et al. (2002) Neurochem. Int. 40:85.
7. Lesort, M. et al. (2000) Prog. Neurobiol. 61:439.

Publications for Transglutaminase 2/TGM2 (4376-TG)(7)

Publications using 4376-TG

• Ninomiya, Y;Matsuda, S;Suzuki, S;Hirata-Tsuchiya, S;Ueda, T;Nakashima, F;Yasuda, K;Shimada, S;Memida, T;Yoshimoto, T;Yamada, S;Ouhara, K;Mizuno, N; Role of transglutaminase 2 in promoting biglycan synthesis in idiopathic gingival fibromatosis BMC oral health 2024-11-21 [PMID: 39574071] (Bioassay, Human)
• LA Farrelly, RE Thompson, S Zhao, AE Lepack, Y Lyu, NV Bhanu, B Zhang, YE Loh, A Ramakrishn, KC Vadodaria, KJ Heard, G Erikson, T Nakadai, RM Bastle, BJ Lukasak, H Zebroski, N Alenina, M Bader, O Berton, RG Roeder, H Molina, FH Gage, L Shen, BA Garcia, H Li, TW Muir, I Maze Histone serotonylation is a permissive modification that enhances TFIID binding to H3K4me3 Nature, 2019-03-13;0(0):. 2019-03-13 [PMID: 30867594] (Bioassay, Human)
• E Png, A Hou, L Tong Mechanistic role of transglutaminase-2 in focal adhesions Sci Rep, 2018-08-17;8(1):12370. 2018-08-17 [PMID: 30120307] (Enzyme Assay, Human)
• C Valero, H Amaveda, M Mora, JM García-Azn Combined experimental and computational characterization of crosslinked collagen-based hydrogels PLoS ONE, 2018-04-17;13(4):e0195820. 2018-04-17 [PMID: 29664953] (Bioassay)
• Addressing proteolytic efficiency in enzymatic degradation therapy for celiac disease Sci Rep, 2016-08-02;6(0):30980. 2016-08-02 [PMID: 27481162] (Bioassay, Plant - Wheat)
• Enzymatically Regulated Peptide Pairing and Catalysis for the Bioanalysis of Extracellular Prometastatic Activities of Functionally Linked Enzymes Sci Rep, 2016-05-03;6(0):25362. 2016-05-03 [PMID: 27140831] (Enzyme Assay, Human)
• Arjomandi M, Frelinger J, Donde A Secreted osteopontin is highly polymerized in human airways and fragmented in asthmatic airway secretions. PLoS ONE, 2011-10-21;6(10):e25678. 2011-10-21 [PMID: 22031818] (Bioassay, Human)

Bioinformatics

• Gene Symbol: TGM2
• Uniprot:
  • P21980()