Recombinant Human GGT5 Protein, CF

R&D Systems | Catalog # 7209-GT

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human GGT5 Protein (7209-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

Sf 21 (baculovirus)

Accession Number

P36269

Applications

Enzyme Activity
View all GGT5 Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human GGT5 protein
Ser30-Tyr586, with an N-terminal 6-His tag.

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

His and Thr388

Predicted Molecular Mass

60 kDa

SDS-PAGE

24-27(mature), 41-46 (mature) & 64-66 (pro) kDa, reducing conditions

Activity

Measured by its ability to hydrolyze glutathione to Glu and Cys-Gly.
The specific activity is >265 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7209-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: GGT5

Gamma-glutamyl transpeptidase-5 (GGT5) is expressed as a pro form, which undergoes autocatalytic cleavage in the endoplasmic reticulum to become the two chain mature form (1). The mature form of GGT is a membrane protein embedded into the outer surface of the plasma membrane through its N-terminal transmembrane domain. It cleaves the gamma-glutamyl peptide bond of glutathione (GSH) and glutathione S-conjugates and transfers the g-glutamyl moiety to acceptors (2). By breaking down extracellular GSH into its constitutive amino acids, its activity is critical for maintaining GSH and cysteine homeostasis. GGT1 and GGT5 are the most fully characterized members of the GGT family (2). Elevated activity of GGT1 in blood has been used to measure liver damage such as that found in hepatocellular carcinoma or alcoholic liver (3). GGT5, also known as g-glutamyl leukotrienase (GGL), may play an important role in leukotriene D4 synthesis and in inflammatory processes.

References

  1. Kinlough C.L. et al. (2005) Methods Enzymol. 401:426.
  2. Heisterkamp N. et al. (2008) Hum. Genet. 123:321.
  3. Whitfield J.B. (2001) Crit. Rev. Clin. Lab. Sci. 38:263.
  4. Han B. et al. (2002) Am. J. Path. 161:481.

Long Name

gamma-Glutamyltransferase 5

Alternate Names

GGT-rel, GGTLA1

Entrez Gene IDs

2687 (Human); 23887 (Mouse)

Gene Symbol

GGT5

UniProt

P36269 (Human)

Additional GGT5 Products

Product Documents for Recombinant Human GGT5 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 GGT5 Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd. Tween is a registered trademark of ICI Americas.

For research use only

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Protocols

View specific protocols for Recombinant Human GGT5 Protein, CF (7209-GT):

Materials
  • Assay Buffer: 50 mM Tris, 0.05% Tween® 20, pH 8.0
  • Recombinant Human GGT5 (rhGGT5) (Catalog # 7209-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 Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhGGT5 to 1 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 50 µL rhGGT5 into wells of plate. Include 50 µL Assay Buffer for a Substrate Blank.
  4. Start reaction by adding 50 µL substrate mixture to all wells.
  5. Read the plate at excitation and emission wavelengths of 540 and 585 nm, respectively, for five minutes in kinetic mode.
  6. 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:

  • rhGGT5: 0.050 µg
  • GSH: 1 mM
  • GIDH: 5 U
  • beta -NAD: 1 mM
  • rhNQO-1: 0.2 μg
  • Resazurin: 0.02 mM

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