Deep mutational scanning reveals the structural basis for α-synuclein activity.
- Author(s): Newberry, Robert W
- Leong, Jaime T
- Chow, Eric D
- Kampmann, Martin
- DeGrado, William F
- et al.
Published Web Locationhttps://doi.org/10.1038/s41589-020-0480-6
Defining the biologically active structures of proteins in their cellular environments remains challenging for proteins with multiple conformations and functions, where only a minor conformer might be associated with a given function. Here, we use deep mutational scanning to probe the structure and dynamics of α-synuclein, a protein known to adopt disordered, helical and amyloid conformations. We examined the effects of 2,600 single-residue substitutions on the ability of intracellularly expressed α-synuclein to slow the growth of yeast. Computational analysis of the data showed that the conformation responsible for this phenotype is a long, uninterrupted, amphiphilic helix with increasing dynamics toward the C terminus. Deep mutational scanning can therefore determine biologically active conformations in cellular environments, even for a highly dynamic multi-conformational protein.