Live-Cell Protein Modification by Boronate-Assisted Hydroxamic Acid Catalysis
Live-Cell Protein Modification by Boronate-Assisted Hydroxamic Acid Catalysis
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DOI：
10.1021/jacs.1c07060
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发表时间：
2021-09-10
影响因子：
15
通讯作者：
Kanai, Motomu
中科院分区：
文献类型：
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作者：
Adamson, Christopher;Kajino, Hidetoshi;Kanai, Motomu
在活细胞内引入蛋白质翻译后修饰（PTM）的选择性方法已被证明对于询问其生物学功能是有价值的。与酶促方法相比，非生物催化应该提供对不同的和新的自然PTM的访问。在此，我们报告的硼酸盐辅助异羟肟酸（BAHA）催化剂系统，它包括一个蛋白质配体，异羟肟酸刘易斯碱，和二醇部分。与含硼酸的酰基供体一致，我们的催化剂利用局部摩尔浓度效应促进酰基转移到目标赖氨酸残基。我们的催化剂系统采用微摩尔试剂浓度，并提供最小的脱靶蛋白反应性。至关重要的是，BAHA对谷胱甘肽有抗性，谷胱甘肽是一种代谢物，它阻碍了许多在活细胞内进行非生物化学的努力。为了展示这种方法，我们在E.大肠杆菌二氢叶酸还原酶在人类细胞中的表达。我们的研究结果进一步建立了著名的硼酸-二醇络合作为一个良好的生物正交反应，在化学生物学和细胞内催化的应用。
Selective methods for introducing protein post-translational modifications (PTMs) within living cells have proven valuable for interrogating their biological function. In contrast to enzymatic methods, abiotic catalysis should offer access to diverse and new-to-nature PTMs. Herein, we report the boronate-assisted hydroxamic acid (BAHA) catalyst system, which comprises a protein ligand, a hydroxamic acid Lewis base, and a diol moiety. In concert with a boronic acid-bearing acyl donor, our catalyst leverages a local molarity effect to promote acyl transfer to a target lysine residue. Our catalyst system employs micromolar reagent concentrations and affords minimal off-target protein reactivity. Critically, BAHA is resistant to glutathione, a metabolite which has hampered many efforts toward abiotic chemistry within living cells. To showcase this methodology, we installed a variety of acyl groups in E. coli dihydrofolate reductase expressed within human cells. Our results further establish the well-known boronic acid-diol complexation as a bona f ide bio-orthogonal reaction with applications in chemical biology and in-cell catalysis.