Max Planck Institute for Developmental Biology, Tübingen, Germany.
University Hospital Tübingen, Division of Translational Oncology, Department of Hematology, Oncology, Clinical Immunology and Rheumatology, University Hospital Tübingen, Germany.
PLoS Biol. 2020 Dec 22;18(12):e3000919. doi: 10.1371/journal.pbio.3000919. eCollection 2020 Dec.
Computational protein design is rapidly becoming more powerful, and improving the accuracy of computational methods would greatly streamline protein engineering by eliminating the need for empirical optimization in the laboratory. In this work, we set out to design novel granulopoietic agents using a rescaffolding strategy with the goal of achieving simpler and more stable proteins. All of the 4 experimentally tested designs were folded, monomeric, and stable, while the 2 determined structures agreed with the design models within less than 2.5 Å. Despite the lack of significant topological or sequence similarity to their natural granulopoietic counterpart, 2 designs bound to the granulocyte colony-stimulating factor (G-CSF) receptor and exhibited potent, but delayed, in vitro proliferative activity in a G-CSF-dependent cell line. Interestingly, the designs also induced proliferation and differentiation of primary human hematopoietic stem cells into mature granulocytes, highlighting the utility of our approach to develop highly active therapeutic leads purely based on computational design.
计算蛋白质设计正在迅速变得更加强大，提高计算方法的准确性将通过消除实验室中经验优化的需要，极大地简化蛋白质工程。在这项工作中，我们旨在使用重支架构策略设计新型粒细胞生成剂，目标是实现更简单和更稳定的蛋白质。所有 4 个经过实验测试的设计都折叠成单体并且稳定，而 2 个确定的结构与设计模型的差异小于 2.5 Å。尽管缺乏与天然粒细胞生成剂显著的拓扑或序列相似性，但 2 个设计与粒细胞集落刺激因子 (G-CSF) 受体结合，并在 G-CSF 依赖性细胞系中表现出强大但延迟的体外增殖活性。有趣的是，这些设计还诱导原代人造血干细胞增殖和分化为成熟粒细胞，突出了我们仅基于计算设计开发高活性治疗先导物的方法的实用性。
