Cloned (Comment) | Organism |
---|---|
recombinant expression of non-glycosylated and glycosylated His6-tagged C4ST-1 construct in Escherichia coli strain BL21 Star(DE3), recombinant expression in Pichia pastoris strain ATCC 76273, both in its non-glycosylated and glycosylated forms, leads to an unstable enzyme. Glycosylation of the yeast-expressed sulfotransferase does not improve its overall stability. The activity of the purified Pichia pastoris expressing C4ST-1 is completely lost after 72 h | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | two-substrate Michaelis-Menten kinetic analysis | Homo sapiens | |
0.00125 | - |
3'-phosphoadenylyl sulfate | pH 7.0, 37°C, recombinant enzyme expressed in Escherichia coli | Homo sapiens | |
0.0038 | - |
chondroitin | pH 7.0, 37°C, recombinant enzyme expressed in Escherichia coli | Homo sapiens | |
0.036 | - |
3'-phosphoadenylyl sulfate | pH 7.0, 37°C, recombinant enzyme expressed in Pichia pastoris | Homo sapiens | |
0.157 | - |
chondroitin | pH 7.0, 37°C, recombinant enzyme expressed in Pichia pastoris | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3'-phosphoadenylyl sulfate + chondroitin | Homo sapiens | - |
adenosine 3',5'-bisphosphate + chondroitin 4'-sulfate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q9NPF2 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | four potential glycosylation sites in human C4ST-1: Asn205, Asn223, Asn321, and Asn342. Enzyme C4ST-1 requires glycosylation for its activity. The enzyme completely loses activity when deglycosylated by treatment with PNGase F. In particular, the glycosylation site Asn342 near the C-terminus is believed to be essential to the activity of C4ST-1 since the mutant devoid of this site lost significant activity compared to the wild-type enzyme. Asn223 and Asn342 are thought to contribute mainly to the stability of the protein rather than to the production of the active enzyme. The recombinant glycosylated form of C4ST-1 shows lower stability than the non-glycosylated form | Homo sapiens |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged C4ST-1 from Escherichia coli strain BL21 Star(DE3) by nickel affinity chromatography | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
bone marrow | - |
Homo sapiens | - |
brain | - |
Homo sapiens | - |
heart | - |
Homo sapiens | - |
leukocyte | from peripheral blood | Homo sapiens | - |
lung | - |
Homo sapiens | - |
lymph node | - |
Homo sapiens | - |
additional information | C4ST-1 is predominantly expressed in spleen, thymus, bone marrow, peripheral blood leukocytes, lymph node, heart, brain, lung, and placenta | Homo sapiens | - |
placenta | - |
Homo sapiens | - |
spleen | - |
Homo sapiens | - |
thymus | - |
Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3'-phosphoadenylyl sulfate + chondroitin | - |
Homo sapiens | adenosine 3',5'-bisphosphate + chondroitin 4'-sulfate | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x 50000, recombinant His6-tagged C4ST-1, SDS-PAGE | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
C4ST-1 | - |
Homo sapiens |
chondroitin-4-O-sulfotransferase | - |
Homo sapiens |
chondroitin-4-sulfotransferase | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
evolution | there are three major C4ST isoforms in Homo sapiens, C4ST-1, C4ST-2, and C4ST-3. Based on a general amino acid sequence analysis, C4STs contain a single cytoplasmic domain followed by helical transmembrane domain and a lumenal domain. C4ST-1 has two additional variants, and both are capable of sulfating unsulfated chondroitin and dermatan with a preference of D-glucuronic acid (GlcA)->GalNAc over L-iduronic acid (IdoA)->GalNAc. C4ST-2 has a lower activity towards unsulfated dermatan than does C4ST-1 | Homo sapiens |
malfunction | even though C4ST-1 and C4ST-2 exhibit broad and overlapping mRNA expression patterns, indicating the functional redundancy of these enzymes, a deficiency or experimental knockdown of C4ST-1 results in a dramatic decrease of cellular and whole-body levels of chondroitin 4'-sulfate. The enzyme completely loses activity when deglycosylated by treatment with PNGase F | Homo sapiens |
metabolism | C4ST-1 is believed to play a distinct regulatory role not only in CS 4-O-sulfation but also in the amount of CS synthesis since none of the other family member C4ST-2 and C4ST-3 can compensate for the loss of C4ST-1 | Homo sapiens |
physiological function | chondroitin sulfates are linear sulfated polysaccharides called glycosaminoglycans. They are important nutraceutical and pharmaceutical products that are biosynthesized through the action of chondroitin sulfotransferases on either an unsulfated chondroitin or a dermatan polysaccharide precursor. The major chondroitin-4-sulfotransferase in Homo sapiens is C4ST-1 | Homo sapiens |