Recombinant Human Sulfamidase/SGSH Protein, CF (Catalog # 8380-SU) hydrolyzes N-linked sulfate group from the non-reducing terminal glucosaminide residues of heparan sulfate.
2 μg/lane of Recombinant Human Sulfamidase/SGSH Protein (Catalog # 8380-SU) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands ...read more
Recombinant Human Sulfamidase/SGSH Protein, CF Summary
Details of Functionality
Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is >0.125 pmol/min/μg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Sulfamidase/SGSH protein Arg23-Leu502 with a C-terminal 6-His tag
>90%, 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
55 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
55-63 kDa, under reducing conditions.
Publications
Read Publication using 8380-SU in the following applications:
Combine 150 µL of 40 µg/mL of rhSGSH and 150 µL of 20 mM of PNCS. Include a Substrate Blank containing 150 µL of Assay Buffer and 150 µL of 20 mM of PNCS.
Incubate at 37 °C for 72 hours.
Stop reactions and Substrate Blank by adding 300 µL of 0.2 M NaOH.
Load into a plate 200 µL from each stopped reaction and Substrate Blank.
Read plate at 510 nm (absorbance) in endpoint mode.
Calculate specific activity:
Specific Activity (pmol/min/µg) =
Adjusted Abs* (OD) x Conversion Factor** (pmol/OD)
Incubation time (min) x amount of enzyme (µg)
*Adjusted for Substrate Blank **Derived using calibration standard 4-Nitrocatechol (PNC).
Per Well:
rhSGSH: 2 µg
Substrate: 5 mM
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human Sulfamidase/SGSH Protein, CF
heparan sulfate sulfatase
HSS
HSSN-sulphoglucosamine sulphohydrolase
MPS3A
MPS3Asulphamidase
N-sulfoglucosamine sulfohydrolase
SFMD
SFMDEC 3.10.1.1
SGSH
Sulfamidase
Sulfoglucosamine sulfamidase
Sulphamidase
Background
Also known as N-sulfoglucosamine sulfohydrolase and heparan N-sulfatase, Sulfamidase/SGSH is an important member of the sulfatase family involved in the degradation of heparan sulfate (HS) (1). Different from the HS specific endosulfatases that remove sulfate from internal GlcNAc residues (2), SGSH removes sulfate group from the non-reducing end glucosamine residues on HS. The SGSH deficiency results in mucopolysaccharidosis type IIIA (MPS IIIA, Sanfilippo A syndrome), an autosomal recessive lysosomal storage disease characterized by neurological dysfunction but relatively mild somatic manifestations (3). Human SGSH shows 88.6% sequence identity with that of mouse sequence.
DiezRoux, G. and Ballabio, A. et al. (2005) Annu. Rev. Genomics Hum. Genet. 6:355.
Morimoto-Tomita, M. et al. (2002) J. Biol. Chem. 277:49175.
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