A target-specific approach for the identification of tyrosine-sulfated hemostatic proteins.
A target-specific approach for the identification of tyrosine-sulfated hemostatic proteins.
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用于鉴定酪氨酸硫酸化止血蛋白的靶标特异性方法。
DOI：
10.1016/j.ab.2009.04.002
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发表时间：
2009
影响因子：
2.9
通讯作者：
Liu,Ming-Cheh
中科院分区：
文献类型：
--
作者：
Liu,Tzu-An;Yasuda,Shin;Williams,FrederickE;Liu,Ming-Yih;Suiko,Masahito;Sakakibara,Yoichi;Yang,Yuh-Shyong;Liu,Ming-Cheh
开发了一种简单的方法来鉴定经过翻译后酪氨酸硫酸化的止血蛋白。该程序包括使用 Sulfinator 预测算法对三个止血途径的成员进行序列分析，然后对培养的 HepG2 人肝癌细胞进行 [35S] 硫酸盐标记，对目标 [35S] 硫酸盐标记的止血蛋白进行免疫沉淀，并对免疫沉淀蛋白进行酪氨酸 O-[35S]硫酸盐分析。鉴定出三种新的酪氨酸硫酸化止血蛋白——蛋白 S、前激肽释放酶和纤溶酶原。这种针对特定目标的方法将允许研究其他生化/生理途径/过程的酪氨酸硫酸化蛋白质，并有助于更好地理解翻译后酪氨酸硫酸化的功能作用。
A simple methodology for the identification of hemostatic proteins that are subjected to posttranslational tyrosine sulfation was developed. The procedure involves sequence analysis of members of the three hemostatic pathways using the Sulfinator prediction algorithm, followed by [35S]sulfate labeling of cultured HepG2 human hepatoma cells, immunoprecipitation of targeted [35S]sulfate-labeled hemostatic proteins, and tyrosine O-[35S]sulfate analysis of immunoprecipitated proteins. Three new tyrosine-sulfated hemostatic proteins—protein S, prekallikrein, and plasminogen—were identified. Such a target-specific approach will allow investigation of tyrosine-sulfated proteins of other biochemical/physiological pathways/processes and contribute to a better understanding of the functional role of posttranslational tyrosine sulfation.