標題: High-Throughput Screening of Sulfated Proteins by Using a Genome-Wide Proteome Microarray and Protein Tyrosine Sulfation System
作者: Huang, Bo-Yu
Chen, Po-Chung
Chen, Bo-Han
Wang, Chen-Chu
Liu, Hsuan-Fu
Chen, Yi-Zao
Chen, Chien-Sheng
Yang, Yuh-Shyong
生物科技學系
Department of Biological Science and Technology
公開日期: 21-Mar-2017
摘要: 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. coil 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.
URI: http://dx.doi.org/10.1021/acs.analchem.6b02853
http://hdl.handle.net/11536/145120
ISSN: 0003-2700
DOI: 10.1021/acs.analchem.6b02853
期刊: ANALYTICAL CHEMISTRY
Volume: 89
起始頁: 3278
結束頁: 3284
Appears in Collections:Articles