Wang Changliu, Zhao Zhenguang, Ghadir Reem, Yang Dechun, Zhang Zhenjia, Ding Zhe, Cao Yuan, Li Yuqing, Fassler Rosi, Reichmann Dana, Zhang Yujie, Zhao Yongli, Liu Can, Bi Xiaobao, Metanis Norman, Zhao Junfeng
Affiliated Cancer Hospital, Guangdong Provincial Key Laboratory of Major Obstetric Diseases, School of Pharmaceutical Sciences, Guangzhou Medical University, Guangzhou 511436, Guangdong P. R. China.
National Research Center for Carbohydrate Synthesis, College of Chemistry and Chemical Engineering, Jiangxi Normal University, Nanchang 330022, Jiangxi P. R. China.
ACS Cent Sci. 2024 Aug 21;10(9):1742-1754. doi: 10.1021/acscentsci.4c01148. eCollection 2024 Sep 25.
Efficient functionalization of peptides and proteins has widespread applications in chemical biology and drug discovery. However, the chemoselective and site-selective modification of proteins remains a daunting task. Herein, a highly efficient chemo-, regio-, and stereoselective hydrosulfuration of ynamide was identified as an efficient method for the precise modification of peptides and proteins by uniquely targeting the thiol group of cysteine (Cys) residues. This novel method could be facilely operated in aqueous buffer and was fully compatible with a wide range of proteins, including small model proteins and large full-length antibodies, without compromising their integrity and functions. Importantly, this reaction provides the -isomer of the corresponding conjugates exclusively with superior stability, offering a precise approach to peptide and protein therapeutics. The potential application of this method in peptide and protein chemical biology was further exemplified by Cys-bioconjugation with a variety of ynamide-bearing functional molecules such as small molecule drugs, fluorescent/affinity tags, and PEG polymers. It also proved efficient in redox proteomic analysis through Cys-alkenylation. Overall, this study provides a novel bioorthogonal tool for Cys-specific functionalization, which will find broad applications in the synthesis of peptide/protein conjugates.
肽和蛋白质的高效功能化在化学生物学和药物发现中具有广泛应用。然而，蛋白质的化学选择性和位点选择性修饰仍然是一项艰巨的任务。在此，烯酰胺的高效化学、区域和立体选择性氢硫化被确定为一种通过独特地靶向半胱氨酸（Cys）残基的硫醇基团来精确修饰肽和蛋白质的有效方法。这种新方法可以在水性缓冲液中轻松操作，并且与广泛的蛋白质完全兼容，包括小型模型蛋白和大型全长抗体，而不会损害它们的完整性和功能。重要的是，该反应仅提供具有优异稳定性的相应共轭物的 - 异构体，为肽和蛋白质治疗提供了一种精确的方法。通过与各种带有烯酰胺的功能分子（如小分子药物、荧光/亲和标签和PEG聚合物）进行Cys生物共轭，进一步例证了该方法在肽和蛋白质化学生物学中的潜在应用。它在通过Cys-烯基化进行氧化还原蛋白质组分析中也被证明是有效的。总体而言，这项研究为Cys特异性功能化提供了一种新的生物正交工具，它将在肽/蛋白质共轭物的合成中找到广泛应用。
