Tyrosine O-sulfation promotes proteolytic processing of progastrin.
Tyrosine O-sulfation is a common post-translational modification of secretory and membrane proteins. The biological function of sulfation is known in only a few proteins, where it appears to enhance protein-protein interactions. Based on known sequences around sulfated tyrosines, a consensus sequence for prediction of target tyrosines has been proposed. However, some proteins are tyrosine sulfated at sites that deviate from the proposed consensus. Among these is progastrin. It is possible that the deviation explains the incomplete sulfation characteristic for bioactive gastrin peptides. In order to test this hypothesis, we have performed site-directed mutagenesis of the gastrin gene followed by heterologous expression in an endocrine cell line. The results show that substitution of the alanyl residue immediately N-terminal to the sulfated tyrosine with an acidic amino acid promotes the sulfation of gastrin peptides. Hence, the study supports the proposed consensus sequence for tyrosine sulfation. Importantly, however, the results also reveal that complete sulfation increases the endoproteolytic maturation of progastrin. Thus, our study suggests an additional function for tyrosine sulfation of possible general significance.
Bundgaard
J R
JR
Department of Clinical Biochemistry, State University Hospital, Copenhagen, Denmark.
Vuust
J
J
Rehfeld
J F
JF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D006224:Cricetinae; D005755:Gastrins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D008239:Lysine / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics; D011499:Protein Processing, Post-Translational; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism
D005755:Gastrins; D011498:Protein Precursors; C094758:gastrin, Asp(11)-; C094760:gastrin, Asp(11)-Phe(12)-; C094759:gastrin, Phe(12)-; C005662:tyrosine O-sulfate; C013706:big gastrin; D014443:Tyrosine; D008239:Lysine