RDM Division of Cardiovascular Medicine, University of Oxford, Oxford, United Kingdom.
Department of Chemistry, University of Oxford, Oxford, United Kingdom.
Sci Rep. 2018 Apr 20;8(1):6333. doi: 10.1038/s41598-018-24568-9.
Both CC and CXC-class chemokines drive inflammatory disease. Tick salivary chemokine-binding proteins (CKBPs), or evasins, specifically bind subsets of CC- or CXC-chemokines, and could precisely target disease-relevant chemokines. Here we have used yeast surface display to identify two tick evasins: a CC-CKBP, P1243 from Amblyomma americanum and a CXC-CKBP, P1156 from Ixodes ricinus. P1243 binds 11 CC-chemokines with K < 10 nM, and 10 CC-chemokines with K between 10 and 100 nM. P1156 binds two ELR + CXC-chemokines with K < 10 nM, and four ELR + CXC-chemokines with K between 10 and 100 nM. Both CKBPs neutralize chemokine activity with IC < 10 nM in cell migration assays. As both CC- and CXC-CKBP activities are desirable in a single agent, we have engineered "two-warhead" CKBPs to create single agents that bind and neutralize subsets of CC and CXC chemokines. These results show that tick evasins can be linked to create non-natural proteins that target subsets of CC and CXC chemokines. We suggest that "two-warhead" evasins, designed by matching the activities of parental evasins to CC and CXC chemokines expressed in disease, would achieve precision targeting of inflammatory disease-relevant chemokines by a single agent.
趋化因子 CC 类和 CXC 类均可引发炎症性疾病。蜱唾液趋化因子结合蛋白（CKBPs），即逃逸蛋白，特异性结合趋化因子 CC 类或 CXC 类亚群，可精确靶向与疾病相关的趋化因子。本研究利用酵母表面展示技术鉴定了两种蜱逃逸蛋白：美洲钝缘蜱的趋化因子 CC 结合蛋白 P1243 和蓖子硬蜱的趋化因子 CXC 结合蛋白 P1156。P1243 以 K < 10 nM 的亲和力结合 11 种 CC 趋化因子，以 K 为 10-100 nM 的亲和力结合 10 种 CC 趋化因子。P1156 以 K < 10 nM 的亲和力结合两种 ELR+CXC 趋化因子，以 K 为 10-100 nM 的亲和力结合四种 ELR+CXC 趋化因子。两种 CKBPs 在细胞迁移实验中以 IC < 10 nM 的浓度中和趋化因子活性。由于单一制剂中同时具有 CC 和 CXC CKBPs 活性是可取的，因此我们构建了“双弹头”CKBPs，以创建能结合和中和趋化因子 CC 和 CXC 亚群的单一制剂。这些结果表明，蜱逃逸蛋白可连接形成非天然蛋白，以靶向趋化因子 CC 和 CXC 亚群。我们建议通过匹配亲本逃逸蛋白与疾病中表达的 CC 和 CXC 趋化因子的活性来设计“双弹头”逃逸蛋白，可通过单一制剂精确靶向与炎症性疾病相关的趋化因子。
