Department of Biochemistry, University of Washington, Seattle, WA 98195, USA.
Institute for Protein Design, University of Washington, Seattle, WA 98195, USA.
Science. 2018 Mar 2;359(6379):1042-1046. doi: 10.1126/science.aaq1739.
The computational design of transmembrane proteins with more than one membrane-spanning region remains a major challenge. We report the design of transmembrane monomers, homodimers, trimers, and tetramers with 76 to 215 residue subunits containing two to four membrane-spanning regions and up to 860 total residues that adopt the target oligomerization state in detergent solution. The designed proteins localize to the plasma membrane in bacteria and in mammalian cells, and magnetic tweezer unfolding experiments in the membrane indicate that they are very stable. Crystal structures of the designed dimer and tetramer-a rocket-shaped structure with a wide cytoplasmic base that funnels into eight transmembrane helices-are very close to the design models. Our results pave the way for the design of multispan membrane proteins with new functions.
具有一个以上跨膜区域的跨膜蛋白的计算设计仍然是一个主要挑战。我们报告了具有 76 到 215 个残基亚基的跨膜单体、同源二聚体、三聚体和四聚体的设计，这些亚基包含两个到四个跨膜区域和多达 860 个总残基，在去污剂溶液中采用目标寡聚化状态。设计的蛋白质在细菌和哺乳动物细胞中定位于质膜，并且在膜中的磁镊展开实验表明它们非常稳定。设计的二聚体和四聚体的晶体结构 - 具有宽阔细胞质基底的火箭形状结构，其漏斗进入八个跨膜螺旋 - 非常接近设计模型。我们的结果为具有新功能的多跨膜蛋白的设计铺平了道路。
