Recombinant B. Thetaiotaomicron Arylsulfatase Protein, CF

R&D Systems | Catalog # 11383-SU

His-tag
R&D Systems
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Key Product Details

  • R&D Systems E. coli-derived Recombinant B. Thetaiotaomicron Arylsulfatase Protein (11383-SU)
  • 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

WP_011107907.1

Applications

Enzyme Activity
View all Bacteroides Thetaiotaomicron Arylsulfatase Proteins and Enzymes »

Product Specifications

Source

E. coli-derived b. thetaiotaomicron Bacteroides Thetaiotaomicron Arylsulfatase protein
Met1-Lys509 (S77C) with a C-terminal 6-His tag

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

Met1 & Leu8

Predicted Molecular Mass

58 kDa & 59 kDa 

SDS-PAGE

54-60 kDa, under reducing conditions

Activity

Measured by its ability to cleave 3'-sulfate from Galactose.
Recombinant Bt.1636 arylsulfatase will cleave >50% Cy5-Fuc labeled sulfo-G2 (Catalog # GL305), as measured under the described conditions.

Scientific Data Images for Recombinant B. Thetaiotaomicron Arylsulfatase Protein, CF

Recombinant Bacteroides Thetaiotaomicron Arylsulfatase His-tag Protein Enzyme Activity Diagram.

Recombinant B. Thetaiotaomicron Arylsulfatase Protein, CF (Catalog # 11383-SU) has specificity for 3S-Galactose. It has been shown to remove 3’-sulfate (3S) from various glycan structures including LacNAc, LNB, Lewis-X and Lewis-A.

Recombinant Bacteroides Thetaiotaomicron Arylsulfatase His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Bacteroides Thetaiotaomicron Arylsulfatase His-tag Protein (Catalog # 11383-SU) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 54-60 kDa under reducing condition.

Recombinant Bacteroides Thetaiotaomicron Arylsulfatase His-tag Protein Enzyme Activity.

Lane 1 contained fluorescent substrate glycan Cy5-Fuc labeled sulfo-G2. Following treatment with Recombinant B. Thetaiotaomicron Arylsulfatase Protein, CF (Catalog # 11383-SU) 3’-sulfate (3S) from galactose was removed and a mobility shift was observed. SDS-PAGE gel was imaged using the red fluorescent channel.

Formulation, Preparation, and Storage

11383-SU
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: Bacteroides Thetaiotaomicron Arylsulfatase

Recombinant Bacteroides thetaiotaomicron Arylsulfatase His-tag, known in the literature as BT1636, is a cell surface-localized member of a diverse sulfatase family involved in mucin degradation (1). BT1636 is active on 3'S-Gal-beta 1,3-GalNAc (core 1) as well as more complex sulfated glycans built around other core structures (1). This broader specificity suggests the enzyme evolved to recognize various linkages and substitutions found in O-glycans. BT1636 displays an alpha / beta / alpha topology with a C-terminal sub-domain, a coordinated calcium ion, a deep pocket for substrates, and an active site with highly conserved residues that interact with the sulfate group (1-3). Bacteroides thetaiotaomicron (Bt) is one of the major bacteria of the human gut microbiome that can utilize O-glycans as a sole source of nutrients and has a requirement for active sulfatases for competitive colonization (4, 5). Although the Bt genome encodes 28 different S1 sulfatases (1), investigation suggests BT1636 is critical as an early enzyme in O-glycan cleavage that allows import and further degradation of O-glycans in the periplasmic space. Bt mutants lacking only sulfatase BT1636 displayed a large defect in colonization competition experiments versus wild-type Bt (1). Given BT1636 is critical to initiation of the complex pathway of mucin degradation, target and inhibition of this enzyme may lead to the ability to inhibit pathways that enable and contribute to disease such as IBD (1). The activity of recombinant BT1636 is demonstrated in an electrophoretic gel mobility shift assay using a fluorophore-labeled glycan as the substrate (6).

References

  1. Luis, A.S. et al. (2021) Nature. 598:332.
  2. Cartmell, A. et al. (2017) Proc. Natl. Acad. Sci. U.S.A. 27:7037.
  3. Ulmer, J.E. et al. (2014) J. Biol. Chem. 289:24289.
  4. Martens, E.C. et al. (2008) Cell Host Microbe 4:447.
  5. Benjdia, A. et al. (2011) J. Biol. Chem. 286:25973.
  6. Wu, Z.L. et al. (2020) Glycobiology 30:970.

Long Name

Bacteroides Thetaiotaomicron Arylsulfatase

Alternate Names

BT1636

Entrez Gene IDs

60927621 (Bacterial)

Gene Symbol

BT_1636

UniProt

WP_011107907.1

Additional Bacteroides Thetaiotaomicron Arylsulfatase Products

Product Documents for Recombinant B. Thetaiotaomicron Arylsulfatase Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant B. Thetaiotaomicron Arylsulfatase Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant B. Thetaiotaomicron Arylsulfatase Protein, CF (11383-SU):

Materials
  • Assay Buffer: 50 mM MES, 10 mM CaCl2, pH 5.5
  • Recombinant Bt.1636 arylsulfatase (rBt. 1636 arylsulfatase) (Catalog # 11383-SU)
  • Cy5-Fuc labeled sulfo-G2 (Cy5-sulfoG2) (Catalog # GL305)
  • 15% SDS-PAGE gel
  • Gel loading dye
  • Gel Imager with Cy5 fluorescent dye detection capability
  1. Dilute rBt. 1636 arylsulfatase to 0.05 µg/µL with Assay Buffer.
  2. Dilute Cy5-sulfoG2 to 0.02 uM with Assay Buffer.
  3. For reaction, combine 10 µL of rBt. 1636 arylsulfatase and 10 µL of Cy5-sulfoG2. For a control, combine 10 µL of Assay Buffer and 10 µL Cy5-sulfoG2.
  4. Incubate reaction and control at 37 °C for 1 hour.
  5. Add Gel loading dye to each tube.
  6. Load half the volume of each reaction and control onto a 15% SDS-PAGE gel. Let samples migrate at least 80% down the gel before stopping.
  7. Acquire gel image with a gel imager.
  8. Determine intensities of each band. Calculate percent cleavage of Cy5-sulfoG2:

     % Cleavage = [ 1 - (Intensity of uncleaved Cy5-sulfoG2 ) ] x 100
Intensity of cleaved and uncleaved bands
Per Reaction:
  • rBt. 1636 arylsulfatase: 0.5 µg
  • Cy5-sulfoG2: 0.2 pmol

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