期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2022
卷号：119
期号：30
DOI：10.1073/pnas.2113400119
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Significance
Proteins capable of binding arbitrary small molecules could enable the generation of new biosensors or medicines. While considerable progress has been made in recent years to design proteins from scratch capable of binding asymmetric molecules, little work has been done to facilitate the binding of symmetric molecules. Here, we present a method for generating libraries of C2 symmetric proteins with diverse central cavities that could be functionalized in the future to bind a range of C2 symmetric small molecules for applications such as ligand controllable cell engineering. We show that 31% of our designed proteins fold to the desired quaternary state, when experimentally characterized, and are hyperstable.
Function follows form in biology, and the binding of small molecules requires proteins with pockets that match the shape of the ligand. For design of binding to symmetric ligands, protein homo-oligomers with matching symmetry are advantageous as each protein subunit can make identical interactions with the ligand. Here, we describe a general approach to designing hyperstable C2 symmetric proteins with pockets of diverse size and shape. We first designed repeat proteins that sample a continuum of curvatures but have low helical rise, then docked these into C2 symmetric homodimers to generate an extensive range of C2 symmetric cavities. We used this approach to design thousands of C2 symmetric homodimers, and characterized 101 of them experimentally. Of these, the geometry of 31 were confirmed by small angle X-ray scattering and 2 were shown by crystallographic analyses to be in close agreement with the computational design models. These scaffolds provide a rich set of starting points for binding a wide range of C2 symmetric compounds.
