title: High-Throughput Screening of Sulfated Proteins by Using a Genome-Wide Proteome Microarray and Protein Tyrosine Sulfation System.
abstract: Protein tyrosine sulfation (PTS) is a widespread posttranslational modification that induces intercellular and extracellular responses by regulating protein-protein interactions and enzymatic activity. Although PTS affects numerous physiological and pathological processes, only a small fraction of the total predicted sulfated proteins has been identified to date. Here, we localized the potential sulfation sites of Escherichia coli proteins on a proteome microarray by using a 3'-phosphoadenosine 5'-phosphosulfate (PAPS) synthase-coupled tyrosylprotein sulfotransferase (TPST) catalysis system that involves in situ PAPS generation and TPST catalysis. Among the 4256 E. coli K12 proteins, 875 sulfated proteins were identified using antisulfotyrosine primary and Cy3-labeled antimouse secondary antibodies. Our findings add considerably to the list of potential proteins subjected to tyrosine sulfation. Similar procedures can be applied to identify sulfated proteins in yeast and human proteome microarrays, and we expect such approaches to contribute substantially to the understanding of important human diseases.
authors.affiliation: Department of Biological Science and Technology, National Chiao Tung University , 75 Boai Street, Hsinchu 300, Taiwan.
authors.forename: Bo-Yu
authors.initials: BY
authors.lastname: Huang
authors.affiliation: Graduate Institute of Systems Biology and Bioinformatics, National Central University , 300 Jhongda Road, Jhongli 320, Taiwan.
authors.forename: Po-Chung
authors.initials: PC
authors.lastname: Chen
authors.affiliation: Department of Biological Science and Technology, National Chiao Tung University , 75 Boai Street, Hsinchu 300, Taiwan.
authors.forename: Bo-Han
authors.initials: BH
authors.lastname: Chen
authors.affiliation: Department of Biological Science and Technology, National Chiao Tung University , 75 Boai Street, Hsinchu 300, Taiwan.
authors.forename: Chen-Chu
authors.initials: CC
authors.lastname: Wang
authors.affiliation: Department of Biological Science and Technology, National Chiao Tung University , 75 Boai Street, Hsinchu 300, Taiwan.
authors.forename: Hsuan-Fu
authors.initials: HF
authors.lastname: Liu
authors.affiliation: Department of Biological Science and Technology, National Chiao Tung University , 75 Boai Street, Hsinchu 300, Taiwan.
authors.forename: Yi-Zao
authors.initials: YZ
authors.lastname: Chen
authors.affiliation: Graduate Institute of Systems Biology and Bioinformatics, National Central University , 300 Jhongda Road, Jhongli 320, Taiwan.
authors.forename: Chien-Sheng
authors.initials: CS
authors.lastname: Chen
authors.affiliation: Department of Biological Science and Technology, National Chiao Tung University , 75 Boai Street, Hsinchu 300, Taiwan.
authors.forename: Yuh-Shyong
authors.identifier: 0000-0003-0177-473X
authors.initials: YS
authors.lastname: Yang
mesh_terms: D000818:Animals; D004331:Drosophila melanogaster / Q000201:enzymology; D048168:Escherichia coli K12; D029968:Escherichia coli Proteins / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D057166:High-Throughput Screening Assays; D006801:Humans; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000378:metabolism; D040081:Protein Array Analysis; D020543:Proteome; D013430:Sulfate Adenylyltransferase / Q000302:isolation & purification / Q000378:metabolism; D015238:Sulfotransferases / Q000302:isolation & purification / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000737:chemistry
chemical_list: D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; D020543:Proteome; C005662:tyrosine O-sulfate; D014443:Tyrosine; C022322:PAPS synthetase; D013430:Sulfate Adenylyltransferase; D015238:Sulfotransferases; C039079:protein-tyrosine sulfotransferase