Literature summary for 2.8.2.5 extracted from

He, W.; Zhu, Y.; Shirke, A.; Sun, X.; Liu, J.; Gross, R.A.; Koffas, M.A.G.; Linhardt, R.J.; Li, M.
Expression of chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris (2017), Appl. Microbiol. Biotechnol., 101, 6919-6928 .

Cloned(Commentary)

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

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 [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Homo sapiens	Q9NPF2	-	-

Posttranslational Modification

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 (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged C4ST-1 from Escherichia coli strain BL21 Star(DE3) by nickel affinity chromatography	Homo sapiens

Source Tissue

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 and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
3'-phosphoadenylyl sulfate + chondroitin	-	Homo sapiens	adenosine 3',5'-bisphosphate + chondroitin 4'-sulfate	-	?

Subunits

Subunits	Comment	Organism
?	x 50000, recombinant His6-tagged C4ST-1, SDS-PAGE	Homo sapiens

Synonyms

Synonyms	Comment	Organism
C4ST-1	-	Homo sapiens
chondroitin-4-O-sulfotransferase	-	Homo sapiens
chondroitin-4-sulfotransferase	-	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

General Information

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