Rational engineering of an erythropoietin fusion protein to treat hypoxia
Rational engineering of an erythropoietin fusion protein to treat hypoxia
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DOI：
10.1093/protein/gzab025
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发表时间：
2021-11-01
影响因子：
2.4
通讯作者：
Silver, Pamela A.
中科院分区：
文献类型：
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作者：
Lee, Jungmin;Vernet, Andyna;Silver, Pamela A.
促红细胞生成素增强氧递送并减少缺氧诱导的细胞死亡，但其促血栓形成活性对于促红细胞生成素在治疗缺氧中的使用是有问题的。我们构建了一种融合蛋白，可以刺激红细胞生成和神经保护，而不会触发血小板生成，血小板是血栓形成的标志物。该蛋白由抗血型糖蛋白A纳米抗体和促红细胞生成素突变体（L108 A）组成。该突变降低了促红细胞生成素受体同型二聚体的激活，而后者可诱导红细胞生成和血栓形成，但维持了组织保护信号。纳米抗体元件与血型糖蛋白A的结合拯救红细胞前体上的同型二聚体促红细胞生成素受体活化。在细胞增殖测定中，融合蛋白在10（-14）M下具有活性，从而允许估计信号传导所需的受体-配体复合物的数目。该融合蛋白在体外和小鼠中刺激红系细胞增殖，并在体外显示神经保护活性。我们的促红细胞生成素融合蛋白提出了一种新的分子治疗缺氧。
Erythropoietin enhances oxygen delivery and reduces hypoxia-induced cell death, but its prothrombotic activity is problematic for use of erythropoietin in treating hypoxia. We constructed a fusion protein that stimulates red blood cell production and neuroprotection without triggering platelet production, a marker for thrombosis. The protein consists of an anti-glycophorin A nanobody and an erythropoietin mutant (L108A). The mutation reduces activation of erythropoietin receptor homodimers that induce erythropoiesis and thrombosis, but maintains the tissueprotective signaling. The binding of the nanobody element to glycophorin A rescues homodimeric erythropoietin receptor activation on red blood cell precursors. In a cell proliferation assay, the fusion protein is active at 10(-14) M, allowing an estimate of the number of receptor-ligand complexes needed for signaling. This fusion protein stimulates erythroid cell proliferation in vitro and in mice, and shows neuroprotective activity in vitro. Our erythropoietin fusion protein presents a novel molecule for treating hypoxia.