Design of stimulus-responsive two-state hinge proteins

• 成果类型: Article
• 署名作者: Praetorius, Florian; Leung, Philip J. Y.; Tessmer, Maxx H.; Broerman, Adam; Demakis, Cullen; Dishman, Acacia F.; Pillai, Arvind; Idris, Abbas; Juergens, David; Dauparas, Justas; Li, Xinting; Levine, Paul M.; Lamb, Mila; Ballard, Ryanne K.; Gerben, Stacey R.; Nguyen, Hannah; Kang, Alex; Sankaran, Banumathi; Bera, Asim K.; Volkman, Brian F.; Nivala, Jeff; Stoll, Stefan; Baker, David
• 署名单位: University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; Medical College of Wisconsin; Medical College of Wisconsin; University of Washington; University of Washington Seattle; United States Department of Energy (DOE); Lawrence Berkeley National Laboratory; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; Howard Hughes Medical Institute
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-12090
• DOI: 10.1126/science.adg7731
• 发表日期: 2023-08-18
• 页码: 754-760
• 关键词: de-novo design; energy landscape

摘要

In nature, proteins that switch between two conformations in response to environmental stimuli structurally transduce biochemical information in a manner analogous to how transistors control information flow in computing devices. Designing proteins with two distinct but fully structured conformations is a challenge for protein design as it requires sculpting an energy landscape with two distinct minima. Here we describe the design of hinge proteins that populate one designed state in the absence of ligand and a second designed state in the presence of ligand. X-ray crystallography, electron microscopy, double electron-electron resonance spectroscopy, and binding measurements demonstrate that despite the significant structural differences the two states are designed with atomic level accuracy and that the conformational and binding equilibria are closely coupled.