Lajoie Marc J, Boyken Scott E, Salter Alexander I, Bruffey Jilliane, Rajan Anusha, Langan Robert A, Olshefsky Audrey, Muhunthan Vishaka, Bick Matthew J, Gewe Mesfin, Quijano-Rubio Alfredo, Johnson JayLee, Lenz Garreck, Nguyen Alisha, Pun Suzie, Correnti Colin E, Riddell Stanley R, Baker David
Institute for Protein Design, University of Washington, Seattle, WA, USA.
Department of Biochemistry, University of Washington, Seattle, WA, USA.
Science. 2020 Sep 25;369(6511):1637-1643. doi: 10.1126/science.aba6527. Epub 2020 Aug 20.
Precise cell targeting is challenging because most mammalian cell types lack a single surface marker that distinguishes them from other cells. A solution would be to target cells using specific combinations of proteins present on their surfaces. In this study, we design colocalization-dependent protein switches (Co-LOCKR) that perform AND, OR, and NOT Boolean logic operations. These switches activate through a conformational change only when all conditions are met, generating rapid, transcription-independent responses at single-cell resolution within complex cell populations. We implement AND gates to redirect T cell specificity against tumor cells expressing two surface antigens while avoiding off-target recognition of single-antigen cells, and three-input switches that add NOT or OR logic to avoid or include cells expressing a third antigen. Thus, de novo designed proteins can perform computations on the surface of cells, integrating multiple distinct binding interactions into a single output.
精确的细胞靶向具有挑战性，因为大多数哺乳动物细胞类型缺乏能将它们与其他细胞区分开来的单一表面标志物。一种解决方案是利用细胞表面存在的特定蛋白质组合来靶向细胞。在本研究中，我们设计了依赖共定位的蛋白质开关（Co-LOCKR），其可执行与、或、非布尔逻辑运算。这些开关仅在所有条件都满足时才通过构象变化激活，在复杂细胞群体中以单细胞分辨率产生快速、不依赖转录的反应。我们利用与门重定向T细胞对表达两种表面抗原的肿瘤细胞的特异性，同时避免对单抗原细胞的脱靶识别，还设计了三输入开关，其添加了非或或逻辑以避免或包含表达第三种抗原的细胞。因此，从头设计的蛋白质可在细胞表面进行计算，将多种不同的结合相互作用整合为单一输出。
