Summary
De novo designed backbones can be used to increase the size of proteins for cryo-EM structure determination (1). In one case, a domain designed with RF-diffusion was inserted into an intracellular loop of a G-protein coupled receptor.
Details
Its design needed to proceed in a stepwise manner to avoid the diffused domain from re-entering the membrane.
Figures
Ref (1)
See also
- Antibodies and nanobodies can desymmetrize proteins for cryoEM
- Membrane proteins can be bulked up for cryo-EM by inserting designed domains, including those with predefined epitopes for antibodies
1.
Gao K, Zhang X, Nie J, Meng H, Zhang W, Tian B, et al. De novo design of a fusion protein tool for GPCR research. Proceedings of the National Academy of Sciences. 2025;122(29). Available from: https://doi.org/10.1073/pnas.2422360122