Campaniҫo André, Baran Marcin, Bowie Andrew G, Longley Daniel B, Harrison Timothy, McGouran Joanna F
School of Chemistry, Trinity College Dublin, Dublin, Ireland.
School of Biochemistry and Immunology, Trinity College Dublin, Dublin, Ireland.
Commun Chem. 2025 Jan 29;8(1):25. doi: 10.1038/s42004-025-01412-6.
The thiol-ene reaction between an alkene and a thiol can be exploited for selective labelling of cysteine residues in protein profiling applications. Here, we explore thiol-ene activation in systems from chemical models to complex cellular milieus, using UV, visible wavelength and redox initiators. Initial studies in chemical models required an oxygen-free environment for efficient coupling and showed very poor activation when using a redox initiator. When thiol-ene activation was performed in protein and cell lysate models, all three initiation methods were successful. Faster thiol-ene reaction was observed as the cysteine and alkene were brought into proximity by a binding event prior to activation, leading to quicker adduct formation in the protein model system than the chemical models. Furthermore, in the protein-protein coupling, none of the activators required an oxygen-free environment. Taken together, these observations demonstrate the broad potential for thiol-ene coupling to be used in protein profiling.
烯烃与硫醇之间的硫醇-烯反应可用于蛋白质分析应用中对半胱氨酸残基进行选择性标记。在此，我们利用紫外线、可见光波长和氧化还原引发剂，探索从化学模型到复杂细胞环境等体系中的硫醇-烯活化作用。化学模型的初步研究表明，高效偶联需要无氧环境，且使用氧化还原引发剂时活化效果很差。当在蛋白质和细胞裂解液模型中进行硫醇-烯活化时，所有三种引发方法均取得成功。由于在活化前通过结合事件使半胱氨酸和烯烃靠近，观察到硫醇-烯反应更快，导致蛋白质模型系统中加合物的形成比化学模型更快。此外，在蛋白质-蛋白质偶联中，没有一种活化剂需要无氧环境。综上所述，这些观察结果证明了硫醇-烯偶联在蛋白质分析中具有广泛的应用潜力。
