用于鉴定SAM结合蛋白的光亲和性SAM类似物。
Photoaffinity SAM analogues for the identification of SAM-binding proteins.
作者信息
Wu Xiangyu, Dong Min
机构信息
State Key Laboratory of Synthetic Biology, Frontiers Science Center for Synthetic Biology, School of Chemical Engineering and Technology, Tianjin University Tianjin 300072 China
Haihe Laboratory of Sustainable Chemical Transformations Tianjin 300192 China.
出版信息
Chem Sci. 2025 Jun 3. doi: 10.1039/d5sc03424h.
-Adenosylmethionine (SAM) serves as an important substrate in a variety of biochemical reactions, and it is important to identify unknown SAM-binding proteins to fully understand the biological functions of SAM. Previous studies on SAM-binding proteins used -Adenosylhomocystein (SAH)-analogues, which mainly identified SAM dependent methyltransferases. Here, we developed and validated three SAM photoaffinity probes to label and enrich SAM-binding proteins. These probes efficiently labeled the known SAM-binding protein Dph2 involved in diphthamide biosynthesis from cell lysates. Using these probes, we enriched SAM-binding proteins from the cell lysates of and . In addition, we validated five SAM binders and revealed the SAM cleavage activities of three of them, including the radical SAM enzyme ArsL, which cleaves SAM to generate methylthioadenosine (MTA), and AcnA and EDD84_07545, which generate -adenosyl-l-homocysteine (SAH). Therefore, our SAM-based photoaffinity probes are promising tools for the identification of SAM-binding proteins.
腺苷甲硫氨酸（SAM）在多种生化反应中作为重要底物，识别未知的SAM结合蛋白对于全面理解SAM的生物学功能至关重要。先前对SAM结合蛋白的研究使用腺苷同型半胱氨酸（SAH）类似物，主要鉴定出依赖SAM的甲基转移酶。在此，我们开发并验证了三种SAM光亲和探针，用于标记和富集SAM结合蛋白。这些探针能有效地从细胞裂解物中标记参与白喉酰胺生物合成的已知SAM结合蛋白Dph2。利用这些探针，我们从 和 的细胞裂解物中富集了SAM结合蛋白。此外，我们验证了五种SAM结合蛋白，并揭示了其中三种的SAM裂解活性，包括自由基SAM酶ArsL，它裂解SAM生成甲硫基腺苷（MTA），以及AcnA和EDD84_07545，它们生成腺苷-L-高半胱氨酸（SAH）。因此，我们基于SAM的光亲和探针是识别SAM结合蛋白的有前途的工具。
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