Pham Phuong Ngoc, Zahradník Jiří, Kolářová Lucie, Schneider Bohdan, Fuertes Gustavo
Laboratory of Biomolecular Recognition, Institute of Biotechnology of the Czech Academy of Sciences, Vestec, Czechia.
Faculty of Science, Charles University, Prague, Czechia.
Front Mol Biosci. 2023 Jul 7;10:1214235. doi: 10.3389/fmolb.2023.1214235. eCollection 2023.
Human interleukin 24 (IL-24) is a multifunctional cytokine that represents an important target for autoimmune diseases and cancer. Since the biological functions of IL-24 depend on interactions with membrane receptors, on-demand regulation of the affinity between IL-24 and its cognate partners offers exciting possibilities in basic research and may have applications in therapy. As a proof-of-concept, we developed a strategy based on recombinant soluble protein variants and genetic code expansion technology to photocontrol the binding between IL-24 and one of its receptors, IL-20R2. Screening of non-canonical -nitrobenzyl-tyrosine (NBY) residues introduced at several positions in both partners was done by a combination of biophysical and cell signaling assays. We identified one position for installing NBY, tyrosine70 of IL-20R2, which results in clear impairment of heterocomplex assembly in the dark. Irradiation with 365-nm light leads to decaging and reconstitutes the native tyrosine of the receptor that can then associate with IL-24. Photocaged IL-20R2 may be useful for the spatiotemporal control of the JAK/STAT phosphorylation cascade.
人白细胞介素24（IL - 24）是一种多功能细胞因子，是自身免疫性疾病和癌症的重要靶点。由于IL - 24的生物学功能依赖于与膜受体的相互作用，按需调节IL - 24与其同源伴侣之间的亲和力在基础研究中提供了令人兴奋的可能性，并且可能在治疗中具有应用价值。作为概念验证，我们开发了一种基于重组可溶性蛋白变体和遗传密码扩展技术的策略，以光控IL - 24与其受体之一IL - 20R2之间的结合。通过生物物理和细胞信号测定相结合的方法，对在两个伴侣的多个位置引入的非天然对硝基苄基酪氨酸（NBY）残基进行筛选。我们确定了一个安装NBY的位置，即IL - 20R2的酪氨酸70，这导致在黑暗中异源复合物组装明显受损。用365纳米光照射会导致去笼化并恢复受体的天然酪氨酸，然后该酪氨酸可以与IL - 24结合。光笼化的IL - 20R2可能有助于对JAK/STAT磷酸化级联反应进行时空控制。
