In?vitro display evolution of IL-6R-binding unnatural peptides ribosomally initiated and cyclized with m-(chloromethyl)benzoic acid
In?vitro display evolution of IL-6R-binding unnatural peptides ribosomally initiated and cyclized with m-(chloromethyl)benzoic acid
复制标题
核糖体引发并用间（氯甲基）苯甲酸环化的 IL-6R 结合非天然肽的体外展示进化
DOI：
10.1016/j.bbrc.2020.11.123
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发表时间：
2021
影响因子：
3.1
通讯作者：
Kawakami Takashi
中科院分区：
文献类型：
--
作者：
Takamori Yukio;Ando Takehiro;Fuji Daisuke;Yokoyama Takumi;Yamamoto Mizuki;Kawakami Takashi
多功能细胞因子白细胞介素（IL）-6及其受体（IL-6 R）的相互作用涉及各种疾病，不仅包括类风湿性关节炎等自身免疫性疾病，还包括2019冠状病毒病（COVID-19）中的癌症和细胞因子风暴。在这项研究中，通过指数富集（SELEX）从mRNA展示的非天然肽库中系统进化的配体对人IL-6 R的核糖体启动和环化与间-（氯甲基）苯甲酸（mClPh）通过遗传密码扩展（正义抑制）纳入使用PURE（蛋白质合成使用重组元件）系统。从随机肽库中筛选到一个与IL-6 R胞外区结合的13-mer非天然mClPh环化肽，并对筛选到的13-mer非天然mClPh环化肽进行体外亲和力成熟，通过下一代测序鉴定出两个与IL-6 R结合的非天然mClPh环化肽。由于环化可以增加肽的蛋白酶抗性，因此本研究中发现的新型IL-6 R结合mClPh环化肽具有用于涉及IL-6/IL-6 R信号传导的各种研究、治疗和诊断应用的潜力。
The interaction of the multifunctional cytokine interleukin (IL)-6 and its receptor (IL-6R) is involved in various diseases, including not only autoimmune diseases such as rheumatoid arthritis but also cancer and cytokine storms in coronavirus disease 2019 (COVID-19). In this study, systematic evolution of ligands by exponential enrichment (SELEX) against human IL-6R from mRNA-displayed unnatural peptide library ribosomally initiated and cyclized withm-(chloromethyl)benzoic acid (mClPh) incorporated by genetic code expansion (sense suppression) was performed using the PURE (Protein synthesis Using Recombinant Elements) system. A novel 13-mer unnatural mClPh-cyclized peptide that binds to the extracellular domain of IL-6R was discovered from an extremely diverse random peptide library.In vitroaffinity maturation of IL-6R-binding unnatural mClPh-cyclized peptide from focused libraries was performed, identifying two IL-6R-binding unnatural mClPh-cyclized peptides by next-generation sequencing. Because cyclization can increase the protease resistance of peptides, novel IL-6R-binding mClPh-cyclized peptides discovered in this study have the potential to be used for a variety of research, therapeutic, and diagnostic applications involving IL-6/IL-6R signaling.