title: New consensus features for tyrosine O-sulfation determined by mutational analysis.
abstract: Tyrosine sulfation is an ubiquitous modification of proteins synthesized along the secretory pathway. It enhances protein-protein interactions and may be necessary for the bioactivity of secreted proteins and peptides. To predict tyrosine sulfation, a consensus has been proposed based on sequence comparisons of known substrates and on in vitro studies using synthetic peptides. This consensus predicts the presence of acidic residues on the amino-terminal side of the target tyrosine as the key feature. Using site-directed mutagenesis, we have examined the role of residues neighboring the sulfation site of an intact protein, human progastrin, in vivo. The results show that the charge of the residue in the amino-terminal position (-1) of the tyrosine is critical and can be neutral or acidic, whereas a basic residue abolishes sulfation. In addition, the degree of sulfation is influenced by the residues in positions -2 and -3. Hence, surprisingly a basic residue in position -2 enhances sulfation. Our data suggest a considerably broader range of substrates for the tyrosylprotein sulfotransferase than hitherto assumed and that the tyrosylprotein sulfotransferase is cell-specifically expressed.
authors.affiliation: Department of Clinical Biochemistry, Rigshospitalet, University of Copenhagen, DK-2100 Copenhagen, Denmark. jensbund@rh.dk
authors.forename: J R
authors.initials: JR
authors.lastname: Bundgaard
authors.forename: J
authors.initials: J
authors.lastname: Vuust
authors.forename: J F
authors.initials: JF
authors.lastname: Rehfeld
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002852:Chromatography, Ion Exchange; D006224:Cricetinae; D004252:DNA Mutational Analysis; D005755:Gastrins / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011498:Protein Precursors / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D005755:Gastrins; D011498:Protein Precursors; D013431:Sulfates; C013706:big gastrin; D014443:Tyrosine; C042873:preprogastrin