Detection of Cysteine Sulfenic Acid on E. coli Proteins with a Biotin-Benzoboroxole Probe,ACS Chemical Biology

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Detection of Cysteine Sulfenic Acid on E. coli Proteins with a Biotin-Benzoboroxole Probe
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2023-06-01 , DOI: 10.1021/acschembio.3c00073
Yaxin Niu ₁ , Zhenzhen Chen ₁ , Zhongyao Jiang ₁ , Yanmei Yang ₁ , Guangzhao Liu ₁ , Xiufen Cheng ₁ , Zhenhao Jiang ₁ , Guanglu Zhang ₁ , Lili Tong ₁ , Bo Tang _{1,

2}

Affiliation

S-sulfenylation of cysteine residues on proteins can effectively change protein structures and accordingly regulate their functions in vivo. Investigation of S-sulfenylation in different biological environments is thus vital for a systematic understanding of cellular redox regulation. In this work, a functional probe, biotin-benzoboroxole (Bio-ben), was designed for the detection of cysteine sulfenic acid (Cys-SOH). The performance of Bio-ben was characterized by small-molecule sulfenic acid, protein models, and proteome tests via mass spectra and western blotting. The results showed that Bio-ben was validated for cysteine sulfenic acid on proteins with good capture efficiency even at low concentrations. Compared with commonly used probes such as dimedone, the current probe has significantly shortened labeling time and exhibited comparable sensitivity. The proposed method provides a new approach for exploring S-sulfenylation in the oxidative modification of proteins and is helpful for related biological and clinical applications.

中文翻译：

使用生物素-苯并硼杂环戊烯探针检测大肠杆菌蛋白质上的半胱氨酸磺酸

蛋白质上半胱氨酸残基的S-磺酰化可以有效改变蛋白质结构并相应地调节其体内功能。因此，研究不同生物环境中的 S-磺酰化对于系统地了解细胞氧化还原调节至关重要。在这项工作中，设计了一种功能探针生物素-苯并硼杂环戊烯（Bio-ben）用于检测半胱氨酸磺酸（Cys-SOH）。Bio-ben 的性能通过小分子次磺酸、蛋白质模型以及通过质谱和蛋白质印迹进行的蛋白质组测试来表征。结果表明，Bio-ben 对半胱氨酸磺酸在蛋白质上的捕获效率进行了验证，即使在低浓度下也具有良好的捕获效率。与常用的探针如双甲酮相比，当前的探针显着缩短了标记时间并表现出相当的灵敏度。该方法为探索蛋白质氧化修饰中的S-磺酰化提供了新的途径，有助于相关的生物学和临床应用。

更新日期：2023-06-01

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