Department of Chemistry , University of Cambridge , Lensfield Road , Cambridge CB2 1EW , U.K.
Institut für Molekulare Medizin, Medizinische Fakultät , Martin-Luther-Universität Halle-Wittenberg , Kurt-Mothes-Stra 3a , 06108 Halle , Germany.
Biochemistry. 2018 Oct 30;57(43):6144-6152. doi: 10.1021/acs.biochem.8b00639. Epub 2018 Oct 16.
The field of chemical site-selective modification of proteins has progressed extensively in recent decades to enable protein functionalization for imaging, drug delivery, and functional studies. In this Perspective, we provide detailed insight into an alternative use of site-selective protein chemistry to probe the role(s) of unpaired Cys residues in the structure and function of disease relevant proteins. Phosphatases are important players in the successful infection of pathogenic bacteria, which represent a significant health burden, particularly in multi-drug-resistant strains. Therefore, a strategy for readily probing the key amino acid role(s) in structure and function may facilitate the targeting and inhibition of these virulence factors. With a dehydroalanine-based posttranslational chemical mutagenesis approach, it is possible to reveal hitherto unknown function(s) of noncatalytic Cys residues and confirm their role and interplay in pathogenic bacterial phosphatases. By selectively modifying reactive sulfhydryl side chains in different protein local environments, this posttranslational site-selective chemical mutagenesis approach reveals structural information about binding pockets and regulatory roles of the modified residues, which can be further validated by conventional site-directed mutagenesis. Ultimately, these new binding pockets can serve as templates for enhanced structure-based drug design platforms and aid the development of potent and specific inhibitors.
近几十年来，蛋白质化学位点选择性修饰领域取得了长足进展，能够实现蛋白质功能化，用于成像、药物递送和功能研究。在这篇观点文章中，我们深入详细地探讨了位点选择性蛋白质化学的另一种用途，即探究未配对半胱氨酸残基在疾病相关蛋白质的结构和功能中的作用。磷酸酶是致病细菌成功感染过程中的重要参与者，致病细菌是一个重大的健康负担，尤其是在多重耐药菌株中。因此，一种易于探究关键氨基酸在结构和功能中作用的策略可能有助于靶向和抑制这些毒力因子。通过基于脱氢丙氨酸的翻译后化学诱变方法，有可能揭示非催化半胱氨酸残基迄今未知的功能，并确认它们在致病细菌磷酸酶中的作用和相互作用。通过在不同蛋白质局部环境中选择性修饰反应性巯基侧链，这种翻译后位点选择性化学诱变方法揭示了有关修饰残基的结合口袋和调节作用的结构信息，这可以通过传统的定点诱变进一步验证。最终，这些新的结合口袋可以作为增强基于结构的药物设计平台的模板，并有助于开发强效和特异性抑制剂。
