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Information on EC 3.10.1.1 - N-sulfoglucosamine sulfohydrolase and Organism(s) Homo sapiens and UniProt Accession P51688

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Homo sapiens
UNIPROT: P51688 not found.
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The taxonomic range for the selected organisms is: Homo sapiens
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
sulfamidase, sgsh, sulphamidase, sulphohydrolase, n-sulfoglucosamine sulfohydrolase, heparin sulfamidase, n-sulphoglucosamine, heparan-n-sulfatase, sulphamate sulphohydrolase, heparan-n-sulphatase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heparan-N-sulfatase
-
N-sulfoglucosamine sulfohydrolase
-
sulfamate sulfohydrolase
-
2-desoxy-2-sulphamido-D-glucose sulphamidase
-
-
-
-
2-desoxy-D-glucoside-2-sulphamate sulphohydrolase
-
-
-
-
heparin sulfamidase
-
-
-
-
sulfamidase
-
-
sulfoglucosamine sulfamidase
-
-
-
-
sulphamate sulphohydrolase
-
-
-
-
sulphamidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
N-sulfo-D-glucosamine + H2O = D-glucosamine + sulfate
show the reaction diagram
catalytic reaction mechanism: the active-site formylglycine (FGly70), which is intrinsically reactive, undergoes hydration to form the resting state of the enzyme with a gem-diol group (step 1). Coordination of one of the hydroxyl groups of the gem-diol to a Ca2+ ion facilitates the development of a negative charge on the O atom as its proton is lost to a base. The negatively charged O atom nucleophilically attacks the sulfur centre of the N-linked sulfate group on the glucosamine substrate (step 2), resulting in a covalently bound enzyme-substrate complex with a pentavalent sulfur transition state. An acid (possibly His181) facilitates the cleavage of the S-N bond by protonating the bridging N atom to form an amine leaving group on the N-desulfated substrate, which diffuses away, leaving an O-sulfated enzyme (step 3). Finally, in a step that underlines the importance of the formylglycine residue, another base (His125) deprotonates the second hydroxyl group, resulting in a negatively charged O atom (step 4) that forms a double bond with the C atom as the C-O bond between it and the bridging O atom of the sulfate group breaks, eliminating the sulfate ion and regenerating the formylglycine residue (step 5)
PATHWAY SOURCE
PATHWAYS
-
-, -
SYSTEMATIC NAME
IUBMB Comments
N-sulfo-D-glucosamine sulfohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY hide
37289-41-1
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-methylumbelliferyl-alpha-D-N-sulfoglucosaminide + H2O
4-methylumbelliferyl alpha-D-glucosaminide + sulfate
show the reaction diagram
-
-
-
?
heparan sulfate + H2O
?
show the reaction diagram
-
-
-
?
heparan sulfate + H2O
? + sulfate
show the reaction diagram
-
-
-
?
heparan sulfate + H2O
? + sulfate
show the reaction diagram
heparin + H2O
? + sulfate
show the reaction diagram
O-(alpha-2-sulfamino-2-deoxy-D-glucopyranosyl)-(1-3)-L-idonic acid
O-(alpha-2-amino-2-deoxy-D-glucopyranosyl)-(1-3)-L-idonic acid + sulfate
show the reaction diagram
-
-
-
-
?
tetrasaccharides + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
heparan sulfate + H2O
?
show the reaction diagram
-
-
-
?
heparan sulfate + H2O
? + sulfate
show the reaction diagram
heparin + H2O
? + sulfate
show the reaction diagram
-
-
-
-
?
additional information
?
-
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
phosphate
-
Cu2+
-
1 mM decreases activity to 12% of control activity
SO42-
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.032 - 0.22
heparan sulfate
0.01
heparin
-
-
0.0107 - 0.01225
tetrasaccharides
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1
phosphate
Homo sapiens
pH 6.7, 37°C, recombinant enzyme
5
sulfate
Homo sapiens
pH 6.7, 37°C, recombinant enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.7
assay at
4.4
-
on heparin sulfate
4.9
-
on heparin
5.2
-
recombinant enzyme
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
assay at
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
despite the low sequence identity between the unique N-sulfatase and the group of O-sulfatases, they share a similar overall fold and active-site architecture, including a catalytic formylglycine, a divalent metal-binding site and a sulfate-binding site. A highly conserved lysine in O-sulfatases is replaced in the N-sulfoglucosamine sulfohydrolase by an arginine (Arg282) that is positioned to bind the N-linked sulfate substrate
malfunction
malfunction
additional information
the enzyme shows low structural flexibility. The consensus active site lies in domain 1 in a narrow pocket at the bottom of a surface cleft and close to the end of the first beta-strand, active site structure, overview. Proposed interactions between the terminal N-sulfoglucosamine residue of the substrate with the enzyme in the active site
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
SPHM_HUMAN
502
0
56695
Swiss-Prot
Secretory Pathway (Reliability: 1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
100000
-
gel filtration, fractogel TSK
110000
-
gel filtration, placenta
115000
-
gel filtration, Superose 12
122000
-
gel filtration, Sephacryl S-300
190000
-
gel filtration
54680
-
calculation from cDNA sequence
56000
57000
-
SDS-PAGE, mature enzyme
62000
-
2 * 62000, SDS-PAGE
63000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
recombinant enzyme, crystal structure analysis. Two SGSH monomers associate noncovalently to form a butterfly-shaped homodimer burying approximately 10.3% of the accessible surface area of each subunit
?
-
x * 56000, mature recombinant enzyme, SDS-PAGE, x * 63000, unprocessed recombinant enzyme, SDS-PAGE
dimer
additional information
the monomeric enzyme subunit comprises of two domains, each centred on a beta-sheet: a large N-terminal domain (domain 1) and a smaller C-terminal domain (domain 2), as is typical for the sulfatase fold. There are 14 beta-strands, 13 alpha-helices and six 310-helices (T1-T6) in total
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
Asn41, Asn151, Asn264 and Asn413 are four of the five N-glycosylation sites
glycoprotein
-
-
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant C-terminally His6-tagged, glycosylated full-length enzyme, small unit and large unit crystal forms, X-ray diffraction structure determination and analysis at 2.0-2.4 A resolution, molecular replacement. The enzyme appears to exist as a homodimer in both crystal forms
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G191R
mutation identified in children with mucopolysaccharidosis type IIIA in Germany. Recombinant mutant enzyme shows no activity
L163P
mutation identified in children with mucopolysaccharidosis type IIIA in Germany. Recombinant mutant enzyme shows no activity
R233X
mutation identified in children with mucopolysaccharidosis type IIIA in Germany. Recombinant mutant enzyme shows no activity
R433W
mutation identified in children with mucopolysaccharidosis type IIIA in Germany. Recombinant mutant enzyme shows no activity
R74C
mutation identified in children with mucopolysaccharidosis type IIIA in Germany. Recombinant mutant enzyme shows no activity
S106R
mutation identified in children with mucopolysaccharidosis type IIIA in Germany. Recombinant mutant enzyme shows 3.3% of wild-type activity
Y403del
mutation identified in children with mucopolysaccharidosis type IIIA in Germany. Recombinant mutant enzyme shows no activity
R206P
-
an adult Sanfilippo type A patient with homozygous mutation R206P in the sulfamidase gene shows a mild clinical phenotype characterized essentially by a moderate nonevolving metal retardation, polymorphism R456H is also found
S298P
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4, sodium acetate, pH 5.0, 30% glycerol, 25 mM NaCl, 0.1 mM DTT, 0.3 mg/ml BSA, 3 month
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant C-terminally His6-tagged full-length enzyme from HEK-293 cells by nickel affinity croomatography
SP-Sepharose column chromatography and Sephacryl S-300 gel filtration
apparent homogeneity
-
purification of recombinant enzyme
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in CHO-K1 cells
expression of wild-type and mutant enzymes in BHK or CHO cells
recombinant expression of C-terminally His6-tagged full-length enzyme in HEK-293 cells
DNA and amino acid sequence determination and anaylsis, genotyping of patients with mucopolysaccharidosis type IIIA, overview
-
expression in Chinese hamster ovary cells
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gene SGSH, DNA and amino acid sequence determination and analysis, expression of mutant S298P sulfamidase in BHK cells, rapid degradation is responsible for the low steady state level of the mutant protein. Only small amounts of the S298P sulfamidase are transported to the lysosomes after processing and secretion, most of the mutant sulfamidase exits the endoplasmic reticulum for proteasomal degradation
-
heterologous expression, using a baculovirus system, of the cDNAs corresponding to eight out of 14 mutant alleles present in the patient group of 26 unrelated Spanish patients with mucopolysaccharidosis IIIA. Expression of the following alleles: p.S66W, p.R74H, p.Q85R, p.R206P, p.L386R, p.R433W, p.R433Q, and c.1079delC. Expression of wild-type and mutant sulfamidases in Sf9 insect cells
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
the enzyme is a target for the development of structure-based drug design for the devastating neurodegenerative disorder mucopolysaccharidosis type IIIA or Sanfilippo A syndrome
medicine
diagnostics
-
a fluorimetric SGSH activity assay is commonly used to examine cells of mucopolysaccharidosis type IIIA patients for N-sulfoglucosamine sulfohydrolase activity, modification of the method for brain homogenates and definition of the parameters for assay linearity, overview
medicine
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Kresse, H.
Mucopolysaccharidosis III A (sanfilipo A disease): deficiency of a heparin sulfamidase in skin fibroblasts and leucocytes
Biochem. Biophys. Res. Commun.
54
1111-1118
1973
Homo sapiens, Mammalia
Manually annotated by BRENDA team
Paschke, E.; Kresse, H.
Multiple forms of 2-deoxy-D-glucoside-2-sulphamate sulphohydrolase from human placenta
Biochem. J.
181
677-684
1978
Homo sapiens, Mammalia
Manually annotated by BRENDA team
Mahuran, D.; Clements, P.; Hopwood, J.J.
A rapid four column purification of 2-deoxy-D-glucoside-2-sulphamate sulphohydrolase from human liver
Biochim. Biophys. Acta
757
359-365
1983
Homo sapiens, Mammalia
Manually annotated by BRENDA team
Freeman, C.; Hopwood, J.J.
Human liver sulphamate sulphohydrolase. Determination of native protein and subunit Mr values and influence of substrate aglycone structure on catalytic properties
Biochem. J.
234
83-92
1986
Homo sapiens, Mammalia
Manually annotated by BRENDA team
Bielicki, J.; Hopwood, J.J.; Melville, E.L.; Anson, D.S.
Recombinant human sulphamidase: expression, amplification, purification and characterization
Biochem. J.
329
145-150
1998
Homo sapiens
Manually annotated by BRENDA team
Perkins, K.J; Byers, S.; Yogalingam, G; Weber, B.; Hopwood, J.J.
Expression and characterization of wild typ and mutant recombinant human sulfamidase. Implications for sanfilippo (mucopolysaccharidosis IIIA) syndrome
J. Biol. Chem.
274
37193-37199
1999
Homo sapiens
Manually annotated by BRENDA team
Anson, D.S.; Bielicki, J.
Sulphamidase
Int. J. Biochem. Cell Biol.
31
363-367
1999
Homo sapiens
Manually annotated by BRENDA team
Gabrielli, O.; Coppa, G.V.; Bruni, S.; Villani, G.R.; Pontarelli, G.; di Natale, P.
An adult Sanfilippo type A patient with homozygous mutation R206P in the sulfamidase gene
Am. J. Med. Genet. A
133
85-89
2005
Homo sapiens
Manually annotated by BRENDA team
Muschol, N.; Storch, S.; Ballhausen, D.; Beesley, C.; Westermann, J.C.; Gal, A.; Ullrich, K.; Hopwood, J.J.; Winchester, B.; Braulke, T.
Transport, enzymatic activity, and stability of mutant sulfamidase (SGSH) identified in patients with mucopolysaccharidosis type III A
Hum. Mutat.
23
559-566
2004
Homo sapiens (P51688), Homo sapiens
Manually annotated by BRENDA team
Savas, P.S.; Hemsley, K.M.; Hopwood, J.J.
Intracerebral injection of sulfamidase delays neuropathology in murine MPS-IIIA
Mol. Genet. Metab.
82
273-285
2004
Homo sapiens
Manually annotated by BRENDA team
Montfort, M.; Garrido, E.; Hopwood, J.J.; Grinberg, D.; Chabas, A.; Vilageliu, L.
Expression and functional characterization of human mutant sulfamidase in insect cells
Mol. Genet. Metab.
83
246-251
2004
Homo sapiens
Manually annotated by BRENDA team
Gliddon, B.L.; Hopwood, J.J.
Enzyme-replacement therapy from birth delays the development of behavior and learning problems in mucopolysaccharidosis type IIIA mice
Pediatr. Res.
56
65-72
2004
Homo sapiens
Manually annotated by BRENDA team
Hemsley, K.M.; Beard, H.; King, B.M.; Hopwood, J.J.
Effect of high dose, repeated intra-CSF injection of sulphamidase on neuropathology in MPS IIIA mice
Genes Brain Behav.
7
740-753
2008
Homo sapiens
Manually annotated by BRENDA team
Meyer, A.; Kossow, K.; Gal, A.; Steglich, C.; Muehlhausen, C.; Ullrich, K.; Braulke, T.; Muschol, N.
The mutation p.Ser298Pro in the sulphamidase gene (SGSH) is associated with a slowly progressive clinical phenotype in mucopolysaccharidosis type IIIA (Sanfilippo A syndrome)
Hum. Mutat.
29
770
2008
Homo sapiens
Manually annotated by BRENDA team
Hemsley, K.M.; King, B.; Hopwood, J.J.
Injection of recombinant human sulfamidase into the CSF via the cerebellomedullary cistern in MPS IIIA mice
Mol. Genet. Metab.
90
313-328
2007
Homo sapiens
Manually annotated by BRENDA team
Urayama, A.; Grubb, J.H.; Sly, W.S.; Banks, W.A.
Mannose 6-phosphate receptor-mediated transport of sulfamidase across the blood-brain barrier in the newborn mouse
Mol. Ther.
16
1261-1266
2008
Homo sapiens (P51688), Mus musculus
Manually annotated by BRENDA team
Muschol, N.; Pohl, S.; Meyer, A.; Gal, A.; Ullrich, K.; Braulke, T.
Residual activity and proteasomal degradation of p.Ser298Pro sulfamidase identified in patients with a mild clinical phenotype of Sanfilippo A syndrome
Am. J. Med. Genet. A
155A
1634-1639
2011
Homo sapiens
Manually annotated by BRENDA team
Sidhu, N.S.; Schreiber, K.; Proepper, K.; Becker, S.; Uson, I.; Sheldrick, G.M.; Gaertner, J.; Kraetzner, R.; Steinfeld, R.
Structure of sulfamidase provides insight into the molecular pathology of mucopolysaccharidosis IIIA
Acta Crystallogr. Sect. D
70
1321-1335
2014
Homo sapiens (P51688)
Manually annotated by BRENDA team
Whyte, L.; Hopwood, J.; Hemsley, K.; Lau, A.
Variables influencing fluorimetric N-sulfoglucosamine sulfohydrolase (SGSH) activity measurement in brain homogenates
Mol. Genet. Metab. Rep.
5
60-62
2015
Homo sapiens, Mus musculus, Mus musculus C57BL/6
Manually annotated by BRENDA team
Maccari, F.; Sorrentino, N.C.; Mantovani, V.; Galeotti, F.; Fraldi, A.; Volpi, N.
Glycosaminoglycan levels and structure in a mucopolysaccharidosis IIIA mice and the effect of a highly secreted sulfamidase engineered to cross the blood-brain barrier
Metab. Brain Dis.
32
203-210
2017
Homo sapiens (P51688), Homo sapiens
Manually annotated by BRENDA team
Jones, S.A.; Breen, C.; Heap, F.; Rust, S.; de Ruijter, J.; Tump, E.; Marchal, J.P.; Pan, L.; Qiu, Y.; Chung, J.K.; Nair, N.; Haslett, P.A.; Barbier, A.J.; Wijburg, F.A.
A phase 1/2 study of intrathecal heparan-N-sulfatase in patients with mucopolysaccharidosis IIIA
Mol. Genet. Metab.
118
198-205
2016
Homo sapiens (P51688), Homo sapiens
Manually annotated by BRENDA team
Wijburg, F.A.; Whitley, C.B.; Muenzer, J.; Gasperini, S.; Del Toro, M.; Muschol, N.; Cleary, M.; Sevin, C.; Shapiro, E.; Bhargava, P.; Kerr, D.; Alexanderian, D.
Intrathecal heparan-N-sulfatase in patients with Sanfilippo syndrome type A A phase IIb randomized trial
Mol. Genet. Metab.
126
121-130
2019
Homo sapiens (P51688), Homo sapiens
Manually annotated by BRENDA team