title: Recombinant expression of selectively sulfated proteins in Escherichia coli.
abstract: Although tyrosine sulfation is a post-translational modification widespread across multicellular eukaryotes, its biological functions remain largely unknown. This is in part due to the difficulties of synthesizing selectively sulfated proteins. Here we report the selective incorporation of sulfotyrosine into proteins in bacteria by genetically encoding the modified amino acid in response to the amber nonsense codon TAG. Moreover, we show that this strategy enables direct expression in Escherichia coli of sulfo-hirudin, previously inaccessible through recombinant methods. The affinity of sulfo-hirudin toward human thrombin is enhanced more than tenfold over that of desulfo-hirudin, suggesting that sulfo-hirudin may offer clinical advantages for use as an anticoagulant. This general approach to the biosynthesis of sulfated proteins should facilitate further study and application of tyrosine sulfation.
authors.affiliation: Department of Chemistry and Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA. ccliu@scripps.edu
authors.forename: Chang C
authors.initials: CC
authors.lastname: Liu
authors.forename: Peter G
authors.initials: PG
authors.lastname: Schultz
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D018389:Codon, Nonsense / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006629:Hirudins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry; D013917:Thrombin / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D018389:Codon, Nonsense; D006629:Hirudins; D011994:Recombinant Proteins; C005662:tyrosine O-sulfate; D014443:Tyrosine; D013917:Thrombin; D000604:Amino Acyl-tRNA Synthetases