AlphaFold accurately predicts distinct conformations based on oligomeric state of a de novo designed protein

Using the molecular modeling program Rosetta, we designed a de novo protein, called SEWN0.1, that binds the heterotrimeric G protein Gq. The design is helical, well-folded, and primarily monomeric in solution at a concentration of 10 uM. However, when we solved the crystal structure of SEWN0.1, we observed a dimer in a conformation incompatible with binding Gq. Unintentionally, we had designed a protein that adopts alternate conformations depending on its oligomeric state. Recently, there has been tremendous progress in the field of protein structure prediction as new methods in artificial intelligence have been used to predict structures with high accuracy. We were curious if the structure prediction method AlphaFold could predict the structure of SEWN0.1 and if the prediction depended on oligomeric state. When AlphaFold was used to predict the structure of monomeric SEWN0.1, it produced a model that resembles the Rosetta design model and is compatible with binding Gq, but when used to predict the structure of a dimer, it predicted a conformation that closely resembles the SEWN0.1 crystal structure. AlphaFolds ability to predict multiple conformations for a single protein sequence should be useful for engineering protein switches.