Literature summary for 2.8.2.20 extracted from

Seibert, C.; Sanfiz, A.; Sakmar, T.P.; Veldkamp, C.T.
Preparation and analysis of N-terminal chemokine receptor sulfopeptides using tyrosylprotein sulfotransferase enzymes (2016), Methods Enzymol., 570, 357-388 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene TPST1, cloning of isozyme TPST-1 without the cytoplasmic N-terminus and the transmembrane domain, which are not required for enzymatic activity, und an N-terminal transferring signal peptide followed by a protein C epitope are N-terminally fused to the catalytic domain (residues 25 to 370 of full-length isozyme), transient recombinant expression in HEK293-T cells. Recombinant expression of His6-tagged isozyme TPST-1 in Escherichia coli strain BL21(DE3) in inclusion bodies	Homo sapiens
gene TPST2, cloning of isozyme TPST-2 without the cytoplasmic N-terminus and the transmembrane domain, which are not required for enzymatic activity, und an N-terminal transferring signal peptide followed by a protein C epitope are N-terminally fused to the catalytic domain (residues 25 to 377 of full-length isozyme), transient recombinant expression in HEK293-T cells	Homo sapiens

Cloned (Comment)

Organism

gene TPST1, cloning of isozyme TPST-1 without the cytoplasmic N-terminus and the transmembrane domain, which are not required for enzymatic activity, und an N-terminal transferring signal peptide followed by a protein C epitope are N-terminally fused to the catalytic domain (residues 25 to 370 of full-length isozyme), transient recombinant expression in HEK293-T cells. Recombinant expression of His6-tagged isozyme TPST-1 in Escherichia coli strain BL21(DE3) in inclusion bodies

Homo sapiens

gene TPST2, cloning of isozyme TPST-2 without the cytoplasmic N-terminus and the transmembrane domain, which are not required for enzymatic activity, und an N-terminal transferring signal peptide followed by a protein C epitope are N-terminally fused to the catalytic domain (residues 25 to 377 of full-length isozyme), transient recombinant expression in HEK293-T cells

Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
Golgi membrane	TPSTs are type II transmembrane proteins with a single alpha-helical transmembrane segment that anchors the catalytic domain in the Golgi lumen	Homo sapiens	139	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
3'-phosphoadenylyl sulfate + protein tyrosine	Homo sapiens	-	adenosine 3',5'-bisphosphate + protein tyrosine-O-sulfate	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	O60507	-	-
Homo sapiens	O60704	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant engineered isozyme TPST-1 from HEK293-T cells by solubilization with 1% v/v Triton X-100, anti-protein C affinity chromatography, and ultrafiltration	Homo sapiens
recombinant engineered isozyme TPST-1 from HEK293-T cells by solubilization with 1% v/v Triton X-100, anti-protein C affinity chromatography, and ultrafiltration. Recombinant His6-tagged TPST-1 from Escherichia coli strain BL21(DE3)inclusion bodies after solubilization and refolding by glutathione affinity chromatography, ultrafiltration, cleavage of the tag by Precission protease, and gel filtration	Homo sapiens

Purification (Comment)

Organism

recombinant engineered isozyme TPST-1 from HEK293-T cells by solubilization with 1% v/v Triton X-100, anti-protein C affinity chromatography, and ultrafiltration

Homo sapiens

recombinant engineered isozyme TPST-1 from HEK293-T cells by solubilization with 1% v/v Triton X-100, anti-protein C affinity chromatography, and ultrafiltration. Recombinant His6-tagged TPST-1 from Escherichia coli strain BL21(DE3)inclusion bodies after solubilization and refolding by glutathione affinity chromatography, ultrafiltration, cleavage of the tag by Precission protease, and gel filtration

Homo sapiens

Renatured (Commentary)

Renatured (Comment)	Organism
functional refolding of recombinant GST-tagged TPST-1 expressed in Escherichia coli strain BL21(DE3) in inclusion bodies using 6 M guanidine hydrochloride (GndHCl), glutathione, and n-dodecyl-beta-D-maltoside	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
3'-phosphoadenylyl sulfate + protein tyrosine	-	Homo sapiens	adenosine 3',5'-bisphosphate + protein tyrosine-O-sulfate	-	?
3'-phosphoadenylyl sulfate + PSGL-1 1-15 peptide	QATEYEYLDYDFLPE-NH2	Homo sapiens	adenosine 3',5'-bisphosphate + sulfated PSGL-1 1-15 peptide	-	?
additional information	in vitro sulfation of peptide substrates, analytical and semipreparative-scale peptide sulfation reactions	Homo sapiens	?	-	-
additional information	in vitro sulfation of peptide substrates, analytical and semipreparative-scale peptide sulfation reactions, product analysis by NMR	Homo sapiens	?	-	-

Synonyms

Synonyms	Comment	Organism
TPST	-	Homo sapiens
TPST-1	-	Homo sapiens
TPST-2	-	Homo sapiens
tyrosylprotein 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
7.5	-	assay at	Homo sapiens

General Information

General Information	Comment	Organism
additional information	the cytoplasmic N-terminus and the transmembrane domain are not required for enzymatic activity	Homo sapiens
physiological function	in most chemokine receptors, one or multiple tyrosine residues have been identified within the receptor N-terminal domain that are, at least partially, modified by posttranslational tyrosine sulfation. For example, tyrosine sulfation has been demonstrated for Tyr3, 10, 14, and 15 of CCR5, for Tyr3, 14, and 15 of CCR8, and for Tyr7, 12, and 21 of CXCR4. For several chemokine receptors that tyrosine sulfation is required for optimal interaction with the chemokine ligands, functional analysis of the role of tyrosine sulfation in chemokine receptor and ligand binding and function, overview. Sulfotyrosine peptides corresponding to the chemokine receptor N-termini are valuable tools in structural and functional studies	Homo sapiens