Department of Biological Science and Technology, National Chiao Tung University , 75 Boai Street, Hsinchu 300, Taiwan.
Graduate Institute of Systems Biology and Bioinformatics, National Central University , 300 Jhongda Road, Jhongli 320, Taiwan.
Anal Chem. 2017 Mar 21;89(6):3278-3284. doi: 10.1021/acs.analchem.6b02853. Epub 2017 Mar 2.
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.
蛋白质酪氨酸硫酸化（PTS）是一种广泛存在的翻译后修饰，通过调节蛋白质-蛋白质相互作用和酶活性，诱导细胞内和细胞外反应。尽管 PTS 影响许多生理和病理过程，但迄今为止，只有一小部分总预测的硫酸化蛋白被鉴定出来。在这里，我们使用涉及原位 PAPS 生成和 TPST 催化的 3'-磷酸腺苷 5'-磷酸硫酸（PAPS）合酶偶联的酪氨酸蛋白硫酸转移酶（TPST）催化系统，在蛋白质组微阵列上定位了大肠杆菌蛋白的潜在硫酸化位点。在 4256 种大肠杆菌 K12 蛋白中，使用抗硫酪氨酸一抗和 Cy3 标记的抗小鼠二抗鉴定出 875 种硫酸化蛋白。我们的发现大大增加了潜在的酪氨酸硫酸化蛋白的列表。类似的程序可以应用于鉴定酵母和人类蛋白质组微阵列中的硫酸化蛋白，我们期望这些方法对理解重要的人类疾病有很大的贡献。
