Recombinant Human Plasma alpha-L-Fucosidase/FUCA2 His, CF

Recombinant Human Plasma alpha-L-Fucosidase/FUCA2 His-tag Protein, CF (Catalog # 11405-GH) is involved in the hydrolysis of terminal L-fucose residues linked via alpha -1,2, alpha -1,3, alpha -1,4, or alpha -1,6. For simplicity only alpha -1,6 is displayed.

Recombinant Human Plasma alpha-L-Fucosidase/FUCA2 His-tag Protein, CF (Catalog # 11405-GH) is measured by its ability to cleave a fluorogenic substrate 4-methylumbelliferyl-alpha -L-fucopyranoside.

2 μg/lane of Recombinant Human Plasma alpha-L-Fucosidase/FUCA2 His-tag Protein (Catalog # 11405-GH) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 52-58 kDa, under reducing conditions.

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

Recombinant Human Plasma alpha-L-Fucosidase/FUCA2 His, CF Summary

• Additional Information: His-tag
• Details of Functionality: Measured by its ability to cleave a fluorogenic substrate 4-methylumbelliferyl-alpha -L-fucopyranoside. The specific activity is >550 pmol/min/μg, as measured under the described conditions.
• Source: Chinese Hamster Ovary cell line, CHO-derived human Plasma alpha-L-Fucosidase/FUCA2 protein
  His29-Ile467 with an N-terminal 6-His tag
• Accession #: Q9BTY2.2
• N-terminal Sequence: His
• Protein/Peptide Type: Recombinant Enzymes
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 52 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: 52-58 kDa, under 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: 50 mM Sodium Acetate, pH 4.5
  • Recombinant Human Plasma alpha-L-Fucosidase/FUCA2 His-tag (rhFUCA2) (Catalog # 11405-GH)
  • Substrate: 4-Methylumbelliferyl-alpha -L-fucopyranoside, 50 mM stock in DMSO
  • Black 96 Well Plate
  • Plate Reader with Fluorescence Read Capability
  1. Dilute rhFUCA2 to 5 µg/mL in Assay Buffer.
  2. Dilute Substrate to 1.6 mM in Assay Buffer.
  3. Load into a plate 50 μL of 5 µg/mL rhFUCA2, and start the reaction by adding 50 μL of 1.6 mM Substrate. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of 1.6 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)
  Per Well:
  • rhFUCA2: 0.25 μg
  • Substrate: 0.8 mM

Notes

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

Alternate Names for Recombinant Human Plasma alpha-L-Fucosidase/FUCA2 His, CF

• A-L-Fucosidase 2
• Alpha-L-fucosidase 2
• Alpha-L-fucoside fucohydrolase 2
• DJ20N2.5
• EC 3.2.1.51
• FUCA2
• fucosidase, alpha-L- 2, plasma
• MGC1314
• plasma alpha-L-fucosidase
• Plasma Fucosidase

Background

Plasma alpha-L-Fucosidase, also known as FucA2 (FUCA2), is an enzyme responsible for hydrolyzing alpha-L-fucose (1). FUCA2 and its isoform FUCA1 are human fucosidases of the glycoside hydrolase family 29A (2).  Whereas FUCA2 is secreted and localized to plasma, FUCA1 is expressed in cellular tissues in the lysosome (3). The mature FUCA2 enzyme exists in the native state as a homotetramer and contains a highly conserved eukaryotic alpha fucosidase catalytic N-terminal barrel structure. In comparison to FUCA1, FUCA2 may not contain a catalytic residue in the sixth beta strand of the barrel structure which may suggest differing substrate specificity or catalytic mechanism (2). FUCA2 has specificity for fucose linked via alpha -1,2, alpha -1,3, alpha -1,4, or alpha -1,6 to the reducing end of glycans (4). As fucosylated glycoconjugates play numerous roles in biological events such as development, apoptosis, and the immune response, fucosylated glycoconjugates are involved in the pathology of inflammation, cancer, and cystic fibrosis (5-8). FUCA2 has been implicated to play a role in several human diseases; FUCA2 was found to be upregulated in 24 tumor types and was associated with negative outcomes for several of the cancer types (9). In addition, FUCA2 was found to play a major role in bacterial adhesion in H. pylori-induced diseases like gastric cancer and duodenal ulcers (10). Consequently, FUCA2 may be of use in both diagnostic and therapeutic research. 

1. Van Elsen, A. et al. (1983) Human Genetics 64:235.
2. Intra, J. et al. (2007) Gene 392:34.
3. Fukushima, H. et al. (1985) Proc. Natl. Acad. Sci. USA 82:1262.
4. Johnson, S. et al. (1991) Comp Biochem Physiol B. 99:479.
5. Hiraishi, K. et al. (1993) Glycobiology 3:381.
6. Solter, D. and Knowles, B.B. (1978) Proc. Natl. Acad. Sci. USA 75:5565.
7. Becker, D.J. and Lowe, J.B. (2003) Glycobiology 13:41R.
8. Liu, T.W. et al. (2009) Proc. Natl. Acad. Sci. USA 106:14581.
9. Zhong, A. et al. (2021) Front. Immunol. 12:758648.
10. Liu, T.W. et al. (2009) Proc Natl Acad Sci. USA 105:14581.

Bioinformatics

• Uniprot:
  • Q9BTY2.2()