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Literature summary for 2.8.2.17 extracted from

  • Yusa, A.; Kitajima, K.; Habuchi, O.
    N-linked oligosaccharides on chondroitin 6-sulfotransferase-1 are required for production of the active enzyme, Golgi localization, and sulfotransferase activity toward keratan sulfate (2006), J. Biol. Chem., 281, 20393-20403.
    View publication on PubMed
Search for more literature for 2.8.2.17

Cloned(Commentary)

Cloned (Comment) Organism
gene C6st, expression of wild-type and mutant FLAG-tagged C6ST-1s in COS-7 cells, addition of tunicamycin completely abolishes N-glycosylation of the recombinant enzyme and enzyme activity, overview Mus musculus

Protein Variants

Protein Variants Comment Organism
N250S site-directed mutagenesis, mutation of a putative N-glycosylation site, deletion of the fourth N-glycosylation site from the N-terminus of the enzyme inhibits production of the active C6ST-1, the mutant remains in the endoplasmic reticulum, while all other wild-type and mutant enzymes are located in the Golgi apparatus Mus musculus
N413S site-directed mutagenesis, mutation of a putative N-glycosylation site, deletion of the fourth or sixth N-glycosylation site from the N-terminus of the enzyme inhibits production of the active C6ST-1 Mus musculus
N457S site-directed mutagenesis, mutation of a putative N-glycosylation site, deletion of the fifth N-glycosylation site from the N-terminus of the enzyme results in a marked loss of the keratan sulfate sulfotransferase activity Mus musculus
N63S site-directed mutagenesis, mutation of a putative N-glycosylation site Mus musculus
N74S site-directed mutagenesis, mutation of a putative N-glycosylation site Mus musculus
N96S site-directed mutagenesis, mutation of a putative N-glycosylation site Mus musculus

Inhibitors

Inhibitors Comment Organism Structure
PNGase F deglycosylation inactivates the enzyme Mus musculus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00034
-
 3'-phosphoadenylyl sulfate pH 6.8, 37°C, recombinant mutant N250S, with chondroitin Mus musculus
0.0004
-
 3'-phosphoadenylyl sulfate pH 6.8, 37°C, recombinant mutant N457S, with chondroitin Mus musculus
0.00076
-
 chondroitin pH 6.8, 37°C, recombinant mutant N250S Mus musculus
0.00103
-
 chondroitin pH 6.8, 37°C, recombinant mutant N457S Mus musculus
0.0011
-
 3'-phosphoadenylyl sulfate pH 6.8, 37°C, recombinant wild-type enzyme, with chondroitin Mus musculus
0.00144
-
 chondroitin pH 6.8, 37°C, recombinant mutant N413S Mus musculus
0.00146
-
 chondroitin pH 6.8, 37°C, recombinant wild-type enzyme Mus musculus
0.0029
-
 Keratan sulfate pH 6.8, 37°C, recombinant mutant N413S Mus musculus
0.0029
-
 3'-phosphoadenylyl sulfate pH 6.8, 37°C, recombinant mutant N413S, with keratan sulfate Mus musculus
0.0038
-
 3'-phosphoadenylyl sulfate pH 6.8, 37°C, recombinant wild-type enzyme, with keratan sulfate Mus musculus
0.00417
-
 3'-phosphoadenylyl sulfate pH 6.8, 37°C, recombinant mutant N413S, with chondroitin Mus musculus
0.005
-
 Keratan sulfate pH 6.8, 37°C, recombinant wild-type enzyme Mus musculus

Localization

Localization Comment Organism GeneOntology No. Textmining
Golgi apparatus
-
 Mus musculus 5794
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
54000
-
 x * 67000, glycosylated recombinant enzyme, SDS-PAGE, x * 54000, deglycosylated recombinant enzyme, SDS-PAGE Mus musculus
67000
-
 x * 67000, glycosylated recombinant enzyme, SDS-PAGE, x * 54000, deglycosylated recombinant enzyme, SDS-PAGE Mus musculus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3'-phosphoadenylyl sulfate + chondroitin Mus musculus
-
 adenosine 3',5'-bisphosphate + chondroitin 6-sulfate
-
 ?
3'-phosphoadenylyl sulfate + keratan sulfate Mus musculus
-
 ?
-
 ?

Organism

Organism UniProt Comment Textmining
Mus musculus O88199 gene Chst3, C6st, or Gst0; isozyme C6ST-1
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein the enzyme contains N-linked oligosaccharide side chains, which have essential roles in the generation of the active enzyme and in its keratan sulfate sulfotransferase activity, overview, six potential N-glycosylation sites Mus musculus

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant FLAG-tagged C6ST-1s from COS-7 cells by anti-FLAG-tag immunoaffinity and wheat germ agglutinin affinity chromatography Mus musculus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3'-phosphoadenylyl sulfate + chondroitin
-
 Mus musculus adenosine 3',5'-bisphosphate + chondroitin 6-sulfate
-
 ?
3'-phosphoadenylyl sulfate + keratan sulfate
-
 Mus musculus ?
-
 ?

Subunits

Subunits Comment Organism
? x * 67000, glycosylated recombinant enzyme, SDS-PAGE, x * 54000, deglycosylated recombinant enzyme, SDS-PAGE Mus musculus

Synonyms

Synonyms Comment Organism
C6ST-1
-
 Mus musculus
chondroitin 6-sulfotransferase-1
-
 Mus musculus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Mus musculus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.4
-
 keratan sulfate sulfotransferase activity, assay at Mus musculus
6.8
-
 chondroitin sulfotransferase activity, assay at Mus musculus