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Literature summary for 2.8.2.8 extracted from

  • Sueyoshi, T.; Kakuta, Y.; Pedersen, L.C.; Wall, F.E.; Pedersen, L.G.; Negishi, M.
    A role of Lys614 in the sulfotransferase activity of human heparan sulfate N-deacetylase/N-sulfotransferase (1998), FEBS Lett., 433, 211-214.
    View publication on PubMed
Search for more literature for 2.8.2.8

Cloned(Commentary)

Cloned (Comment) Organism
DNA sequence determination, analysis, and expression of the wild-type and mutated sulfotransferase domain of the bifunctional enzyme in Escherichia coli BL21 cells as fusion tagged protein Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
18 mg/ml, 0.1 M Tris-HCl, pH 8.3, 0.1 M NaCl, 4 mM 3'-phosphoadenosine 5'-phosphate, reservoir solution: Bis-Tris-propane, pH 7.0, 0.2 M NaCl, 30% PEG 3000, room temperature, X-ray diffraction structure determination and analysis Homo sapiens
purified recombinant bacterially expressed sulfotransferase domain Homo sapiens

Protein Variants

Protein Variants Comment Organism
K614A site-directed mutagenesis of the sulfotransferase domain of the bifunctional enzyme, complete loss of N-sulfotransferase activity Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.02
-
 3'-phosphoadenylylsulfate pH 7.0, 37°C Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
 bifunctional enzyme possessing heparan sulfate N-deacetylase and N-sulfotransferase activity
-

Purification (Commentary)

Purification (Comment) Organism
recombinant fusion tagged sulfotransferase domain from Escherichia coli BL21 cells by affinity chromatography Homo sapiens

Reaction

Reaction Comment Organism Reaction ID
3'-phosphoadenylyl sulfate + [heparan sulfate]-glucosamine = adenosine 3',5'-bisphosphate + [heparan sulfate]-N-sulfoglucosamine residues 558-882 provide the sulfotransferase domain of the bifunctional enzyme, with Lys614 as a catalytically important conserved residue Homo sapiens
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
231
-
 purified recombinant sulfotransferase domain, substrate is at all positions completely desulfated heparin Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3'-phosphoadenylyl sulfate + [heparan sulfate]-glucosamine substrate needs to be deacetylated Homo sapiens adenosine 3',5'-bisphosphate + [heparan sulfate]-N-sulfoglucosamine
-
 ?
3'-phosphoadenylylsulfate + N,O-desulfated heparan sulfate no activity with Homo sapiens adenosine 3',5'-bisphosphate + O-desulfated heparan sulfate
-
 ?
3'-phosphoadenylylsulfate + N-acetylated heparan sulfate no activity with Homo sapiens ?
-
 ?
3'-phosphoadenylylsulfate + N-desulfated heparan sulfate best substrate Homo sapiens adenosine 3',5'-bisphosphate + heparan sulfate
-
 ?
additional information no activity with N-acetylated substrates Homo sapiens ?
-
 ?

Synonyms

Synonyms Comment Organism
HSNST
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Homo sapiens