Recombinant Human GGT1 His-tag Protein, CF

R&D Systems | Catalog # 10977-GT

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human GGT1 His-tag Protein (10977-GT)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

CHO

Accession Number

P19440.2

Applications

Enzyme Activity
View all gamma-Glutamyltransferase 1/GGT1 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human gamma-Glutamyltransferase 1/GGT1 protein
Pro27-Tyr569 with a C-terminal 6-His

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Pro27 & Thr381

Predicted Molecular Mass

39 kDa (heavy chain) & 21 kDa (light chain)

SDS-PAGE

56-69 kDa & 22-26 kDa, under reducing conditions

Activity

Measured by its ability to hydrolyze glutathione to Glu and Cys-Gly.
The specific activity is >1300 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human GGT1 His-tag Protein, CF

Recombinant Human GGT1 His-tag Protein Enzyme Activiy.

Recombinant Human GGT1 His-tag (Catalog # 10977-GT) is measured by its ability to hydrolyze glutathione to Glu and Cys-Gly.

Recombinant Human GGT1 His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Human GGT1 His-tag Protein (Catalog # 10977-GT) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 56-69 kDa and 22-26 kDa, under reducing conditions.

Formulation, Preparation, and Storage

10977-GT
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: gamma-Glutamyltransferase 1/GGT1

Gamma-glutamyl transferase 1 (GGT1), also known as Gamma-glutamyl transpeptidase 1, is a ubiquitous enzyme that cleaves the gamma-glutamyl bond substrates where the glutamate moiety is available. This makes GGT1 crucial for metabolism of glutathione, the most important cellular antioxidant in humans and for its role in leukotriene biosynthesis, required for lipid metabolism. Although GGT1 comes from a 7 member multigene family, only GGT1 and GGT5 have enzymatic activity (1, 2). GGT5 has 40% amino acid homology with GGT1 and only 4% of the activity (2, 3) making GGT1 the primary contributor to enzymatic activity in the cells. GGT1 is synthesized as a single 569 amino acid polypeptide that is autocleaved by threonine 381 into a mature, activated, heterodimeric glycoprotein composed of a large subunit with an N-terminal hydrophobic transmembrane domain that anchors the enzyme to the membrane and a small catalytic subunit containing the catalytic nucleophile threonine 381 (4, 5). The human GGT1 active site is located in a deep channel but remains in an open configuration (4). It has a unique buried chloride ion that is integral to the protein structure (4). GGT1 contributes to cysteine homeostasis, intracellular redox, and inflammation (2, 6-8) and has been implicated to play a role in Parkinson and cardiovascular disease (5, 9), is a diagnostic marker of several diseases including liver and pancreatic cancer and is induced in several cancers to contribute to resistance to chemotherapeutic agents (7, 10-14). 

References

  1. Pawlak, A. et al. (1988) J. Biol. Chem. 263:9913.
  2. Wickham, S. et al. (2011) Anal. Biochem. 414:208.
  3. Heisterkamp, N. et al. (1991) Proc. Natl. Acad. Sci. USA 88:6303.
  4. West, M.B. et al. (2013) J Biol. Chem. 288:31902.
  5. Ndrepepa, G. et al. (2018) Clin. Chim. Acta. 476:130.
  6. Lieberman, M.W. et al. (1996) Proc. Natl. Acad. Sci. 93: 7923.
  7. Hanigan, M.H. (2014) Adv. Cancer Res. 122:103.
  8. Corti, A. et al. (2020) Free Radic. Biol. Med.160:807.
  9. Sian, J. et al. (1994) Ann. Neurol. 36:356.
  10. Whitfield, J. B. (2001) Crit. Rev. Clin. Lab Sci. 38:263.
  11. Lyu, S.C. et al. (2021) Cancer Manag. Res. 13:4887.
  12. Carr, B.I. et al. (2021) J. Transl. Sci. 7:10.15761.
  13. Sayan, M. et al. (2021) Wiad. Lek. 74: 1804.
  14. Jia, Y. et al. (2021) J. Oncol. 9120265.

Long Name

gamma-Glutamyltransferase 1

Alternate Names

CD224, D22S672, D22S73, GGT, GGT-1, GGTD

Entrez Gene IDs

2678 (Human)

Gene Symbol

GGT1

UniProt

P19440.2

Additional gamma-Glutamyltransferase 1/GGT1 Products

Product Documents for Recombinant Human GGT1 His-tag Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human GGT1 His-tag Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human GGT1 His-tag Protein, CF (10977-GT):

Materials
  • Assay Buffer: 50 mM Tris, 0.05% Tween® 20, pH 8.0
  • Recombinant Human GGT-1 (rhGGT-1) (Catalog # 10977-GT)
  • Recombinant Human NQO-1 (rhNQO-1) (Catalog # 7567-DH)
  • Glutamate Dehydrogenase (GIDH) (Sigma, Catalog # G7882), 200 U/mL stock in 50 mM Tris, 0.05% Tween® 20, pH 8.0
  • Nicotinamide Adenine Dinucleotide ( beta -NAD) (Sigma, Catalog # N6522), 100 mM stock in deionized water
  • Resazurin (Catalog # AR002)
  • Glutathione, reduced (GSH) (Amresco, Catalog # 399), 250 mM stock in deionized water
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax M5 by Molecular Devices) or equivalent
  1. Dilute rhGGT-1 to 0.4 µg/mL in Assay Buffer.
  2. Prepare substrate mixture containing 2 mM GSH, 100 U/mL GIDH, 2 mM beta -NAD, 40 µM Resazurin and 4 µg/mL rhNQO-1 in Assay Buffer.
  3. Load in a plate 50 µL of 0.4 µg/mL rhGGT-1, and start the reaction by adding 50 µL of substrate mixture. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of substrate mixture.
  4. Read at excitation and emission wavelengths of 540 and 585 nm, respectively, for five minutes in kinetic mode.
  5. Calculate 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 Resorufin (Sigma, Catalog # R3257) 

Per Well:

  • rhGGT-1: 0.020 µg
  • GSH: 1 mM
  • GIDH: 5 U
  • beta -NAD: 1 mM
  • rhNQO-1: 0.2 µg
  • Resazurin: 0.02 mM

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