Detection of tyrosine sulfation on proteins.
Detection of tyrosine sulfation on proteins.
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DOI：
10.1002/0471140864.ps1407s80
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发表时间：
2015-04-01
影响因子：
--
通讯作者：
Al Ubaidi, Muayyad R
中科院分区：
文献类型：
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作者：
Kanan, Yogita;Al Ubaidi, Muayyad R
酪氨酸硫酸化是一种翻译后修饰（PTM），其中硫酸盐基团被添加到酪氨酸部分。这种PTM负责加强蛋白质之间的相互作用。研究这种PTM的一个缺点是缺乏一种可以检测所有酪氨酸磺酸蛋白的抗体。此外，由于硫酸酪氨酸的不稳定性质，除非使用特殊的修饰程序，否则质谱等其他技术无法用于研究该PTM。这需要相当的技能和知识的质谱。本单元描述了一种体外技术，可以通过放射性标记重组蛋白来研究酪氨酸磺酸蛋白。然后对蛋白质进行氢氧化钡水解和薄层电泳（TLE）。在TLE分析之前，放射性硫酸酪氨酸与非放射性硫酸酪氨酸标准物共定位，确定蛋白质为硫酸酪氨酸蛋白。该技术的优点是，它可以毫无偏差地识别所有酪氨酸硫酸化蛋白，是唯一一种识别蛋白质中酪氨酸硫酸残基的技术。
Tyrosine sulfation is a post-translational modification (PTM) where a sulfate group is added to a tyrosine moiety. This PTM is responsible for strengthening interaction between proteins. One of the drawbacks of studying this PTM is the lack of an antibody that can detect all tyrosine-sulfated proteins. In addition, due to the labile nature of the tyrosine sulfate, other techniques such as mass spectrometry cannot be used to study this PTM unless special modification procedures are used. This requires considerable skill and knowledge of mass spectrometry. This unit describes an in vitro technique that can be used to study tyrosine-sulfated proteins by radiolabeling the recombinant protein. The protein is then subject to barium hydroxide hydrolysis and thin-layer electrophoresis (TLE). Co-localization of radioactive tyrosine-sulfate with nonradioactive tyrosine sulfate standard spiked in before TLE analysis identifies a protein as tyrosine-sulfated protein. The advantage of this technique is that, it identifies all tyrosine-sulfated proteins without any bias and is the only technique that identifies the tyrosine sulfate residues in the protein.