Glyoxalase I (also lactoylglutathione lyase, methylglyoxalase, and glx I) is a 21 kDa member of the Glyoxalase I family. The enzyme is an isomerase that catalyzes the formation of S-D-lactoylglutathione from the hemimercaptal adduct that forms spontaneously between methylglyoxal and reduced GSH (1-4). The monomeric subunit for human Glyoxalase I is 184 amino acids (aa) in length. In the mature protein, the methionine at the N-terminus is removed. Human Glyoxalase I exists in three separable isoforms as homo-and hetero-dimers of two allelic subunit variants, which differ in charge (1). The isoforms are formed when residue 19 is changed from cysteine to tyrosine and residue 111 is changed from glutamine to alanine. Each subunit binds one Zn2+ atom (1, 3-4). The protein is made up of multiple beta strands and alpha helical regions. Human Glyoxalase I shares 91% and 90% aa sequence identity with rat and mouse Glyoxalase I, respectively. The enzyme is ubiquitously expressed and is also present in many tumor cell lines, in which its concentration is often upregulated (1). The biological role of the enzyme remains unclear, but the glyoxalase system detoxifies the precursors of advanced glycation end products, which take part in the pathogenesis of vascular, diabetic, and uremic complications (5).
Recombinant Human Glyoxalase I Protein, CF
R&D Systems | Catalog # 4959-GL
Loading...
Key Product Details
- R&D Systems E. coli-derived Recombinant Human Glyoxalase I Protein (4959-GL)
- 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
E. coli
Accession Number
Applications
Enzyme Activity
Loading...
Product Specifications
Source
E. coli-derived human Glyoxalase I protein
Ala2-Met184, with an N-terminal Met and 6-His tag
Ala2-Met184, with an N-terminal Met and 6-His tag
Purity
>85%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Met
Predicted Molecular Mass
22 kDa
SDS-PAGE
25 kDa, reducing conditions
Activity
Measured by its ability to catalyze the formation of S-D-lactoylglutathione from the hemimercaptal adduct that forms spontaneously between methylglyoxal and reduced glutathione.
The specific activity is >100 nmol/min/µg, as measured under the described conditions.
The specific activity is >100 nmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
4959-GL
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris-HCl and DTT. |
| Reconstitution | Reconstitute at 0.5 mg/mL in sterile, deionized water.
Loading...
|
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Calculators
Background: Glyoxalase I
References
- Ridderstrom, M. & B. Mannervik (1996) Biochem. J. 314:463.
- Marmstal, E. & B. Mannervik (1981) FEBS Lett. 131:301.
- Kim, N-S. et al. (1993) J. Biol. Chem. 268:11217.
- Ranganathan, S. et al. (1993) J. Biol. Chem. 268:5661.
- Kalousova, M. et al. (2007) Ann. N. Y. Acad. Sci. 1126:268.
Alternate Names
Aldoketomutase, GLO1, GLOD1, Glx I, GLYI, Methylglyoxalase
Gene Symbol
GLO1
UniProt
Additional Glyoxalase I Products
Product Documents for Recombinant Human Glyoxalase I Protein, CF
Product Specific Notices for Recombinant Human Glyoxalase I Protein, CF
For research use only
Citations for Recombinant Human Glyoxalase I Protein, CF
Powered by Bioz
Customer Reviews for Recombinant Human Glyoxalase I Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human Glyoxalase I Protein, CF and earn rewards!
Have you used Recombinant Human Glyoxalase I Protein, CF?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Protocols
View specific protocols for Recombinant Human Glyoxalase I Protein, CF (4959-GL):
Materials
- Assay Buffer: 0.1 M Sodium Phosphate, pH 7.0
- Recombinant Human Glyoxalase I (rhGlyoxalase I) (Catalog # 4959-GL)
- Glutathione, Reduced (GSH) (Amresco, Catalog # 0399)
- Methylglyoxal solution, 40% (Sigma, Catalog # M0252)
- 96-well Clear UV Plate (Costar, Catalog # 3635)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Prepare 100 mM GSH in deionized water. Note: Prepare fresh.
- Dilute 40% (6.48 M) Methylglyoxal solution to 100 mM in Assay Buffer. Note: Prepare fresh.
- Combine 1420 µL Assay Buffer, 40 µL 100 mM GSH, and 40 µL 100 mM Methylglyoxal to make the Substrate Mixture.
- Incubate at room temperature for 15 minutes.
- Dilute rhGlyoxalase I to 0.4 ng/µL in Assay Buffer.
- Load 50 µL of 0.4 ng/µL rhGlyoxalase I in a plate, and start the reaction by loading 150 µL of Substrate Mixture. Include a Substrate Blank containing 50 µL of Assay Buffer and 150 µL of Substrate Mixture.
- Read at 240 nm (absorbance) in kinetic mode for 5 minutes. Include a 5 second mix before the first read and a 3 second mix between reads.
- Calculate specific activity:
|
Specific Activity (nmol/min/µg) = |
Adjusted Vmax* (OD/min) x Conversion Factor** (nmol/OD) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard S-Lactoylglutathione (Sigma, Catalog # L7140).
Per Well:- rhGlyoxalase I: 0.020 µg
- Glutathione: 2 mM
- Methylglyoxal: 2 mM