title: Slow and bimolecular folding of a de novo designed monomeric protein DS119.
abstract: De novo protein design offers a unique means to test and advance our understanding of how proteins fold. However, most current design methods are native structure eccentric and folding kinetics has rarely been considered in the design process. Here, we show that a de novo designed mini-protein DS119, which folds into a βαβ structure, exhibits unusually slow and concentration-dependent folding kinetics. For example, the folding time for 50 μM of DS119 was estimated to be ~2 s. Stopped-flow fluorescence resonance energy transfer experiments further suggested that its folding was likely facilitated by a transient dimerization process. Taken together, these results highlight the need for consideration of the entire folding energy landscape in de novo protein design and provide evidence suggesting nonnative interactions can play a key role in protein folding.
authors.affiliation: BNLMS, State Key Laboratory for Structural Chemistry of Unstable and Stable Species, Peking-Tsinghua Center for Life Sciences at College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhu
authors.forename: Ziwei
authors.initials: Z
authors.lastname: Dai
authors.forename: Huanhuan
authors.initials: H
authors.lastname: Liang
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.forename: Feng
authors.initials: F
authors.lastname: Gai
authors.forename: Luhua
authors.initials: L
authors.lastname: Lai
mesh_terms: D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008958:Models, Molecular; D009154:Mutation; D015202:Protein Engineering; D017510:Protein Folding; D055503:Protein Multimerization; D058849:Protein Refolding; D020836:Protein Structure, Quaternary; D058767:Protein Unfolding; D011506:Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: D011506:Proteins