Recombinant Human beta-Glucuronidase/GUSB Protein, CF

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

Recombinant Human beta-Glucuronidase/GUSB Protein, CF Summary

• Details of Functionality: Measured by its ability to hydrolyze 4-methylumbelliferyl-beta -D-glucuronide. The specific activity is >4,000 pmol/min/μg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human beta-Glucuronidase/GUSB protein
  Met1-Thr651, with a C-terminal 6-His tag
• Accession #: AAA52561
• N-terminal Sequence: Leu23
• Protein/Peptide Type: Recombinant Enzymes
• Gene: GUSB
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 73 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: 75-85 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris and NaCl.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • Assay Buffer: 100 mM Sodium Acetate, pH 3.5
  • Recombinant Human beta ‑Glucuronidase/GUSB (rhGUSB) (Catalog # 6144-GH)
  • Substrate: 4-Methylumbelliferyl beta -D-glucuronide (Sigma, Catalog # M9130), 50 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhGUSB to 4 ng/μL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load into a plate 50 μL of 4 ng/μL rhGUSB and start the reaction by adding 50 μL of 2 mM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 2 mM Substrate.
  4. Read plate at excitation and emission wavelengths of 365 nm and 445 nm, respectively, in kinetic mode for 5 minutes.
  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 4-Methylumbelliferone (4-MU) (Sigma, Catalog # M1381).

  Per Well:
  • rhGUSB: 0.200 μg
  • Substrate: 1 mM

Notes

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

Alternate Names for Recombinant Human beta-Glucuronidase/GUSB Protein, CF

• beta-D-glucuronidase
• Beta-G1
• beta-glucuronidase
• BG
• EC 3.2.1.31
• FLJ39445
• glucuronidase, beta
• GUSB
• MPS7

Background

Human beta -Glucuronidase (EC 3.2.1.31) encoded by the GUSB gene is a lysosomal hydrolase involved in the stepwise degradation of glucuronic acid-containing glycosaminoglycans that include heparan sulfate, chondroitin sulfate and hyaluronan (1). The enzyme is only active on the glucuronic acid of the non-reducing end. The native protein has been reported as a tetrameric glycoprotein composed of identical subunits (1, 2). Mutations in the GUSB gene are linked to mucopolysaccharidosis type VII (3). Accumulation of partially degraded glycosaminoglycans, with glucuronic acid residues at the non-reducing termini, are usually found in the lysosomes of patients with the disease (3). It has also been reported that this enzyme may contribute to the depletion of chondroitin from cartilage and thereby facilitate the damage of joints in rheumatoid arthritis (4).

1. Shipley, J.M. et al. (1993) Am. J. Hum. Genet. 52:517.
2. Oshima, A, et al. (1987) Proc. Natl. Acad. Sci. USA 84:685.
3. Bell, C.E. Jr. et al. (1977) J. Clin. Invest. 59:97.
4. Ortutay, Z. et al. (2003) Arthritis Rheum. 48:2163.

Publications for beta-Glucuronidase/GUSB (6144-GH)(1)

Publication using 6144-GH

• Lietzan, AD;Simpson, JB;Walton, WG;Jariwala, PB;Xu, Y;Boynton, MH;Liu, J;Redinbo, MR; Microbial ?-glucuronidases drive human periodontal disease etiology Science advances 2023-05-05 [PMID: 37146137] (Bioassay, Human)

Bioinformatics

• Gene Symbol: GUSB
• Uniprot:
  • AAA52561()