title: Structure of a designed tetrahedral protein assembly variant engineered to have improved soluble expression.
abstract: We recently reported the development of a computational method for the design of coassembling multicomponent protein nanomaterials. While four such materials were validated at high-resolution by X-ray crystallography, low yield of soluble protein prevented X-ray structure determination of a fifth designed material, T33-09. Here we report the design and crystal structure of T33-31, a variant of T33-09 with improved soluble yield resulting from redesign efforts focused on mutating solvent-exposed side chains to charged amino acids. The structure is found to match the computational design model with atomic-level accuracy, providing further validation of the design approach and demonstrating a simple and potentially general means of improving the yield of designed protein nanomaterials.
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, 98195.|Graduate Program in Molecular and Cellular Biology, University of Washington, Seattle, Washington, 98195.
authors.forename: Jacob B
authors.initials: JB
authors.lastname: Bale
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, 98195.
authors.forename: Rachel U
authors.initials: RU
authors.lastname: Park
authors.affiliation: Department of Chemistry and Biochemistry, UCLA, Los Angeles, California, 90095.
authors.forename: Yuxi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, 98195.|Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, 20147.
authors.forename: Shane
authors.initials: S
authors.lastname: Gonen
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, 20147.
authors.forename: Tamir
authors.initials: T
authors.lastname: Gonen
authors.affiliation: Institute for Genomics and Proteomics, UCLA-DOE, Los Angeles, California, 90095.
authors.forename: Duilio
authors.initials: D
authors.lastname: Cascio
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, 98195.|Institute for Protein Design, University of Washington, Seattle, Washington, 98195.
authors.forename: Neil P
authors.initials: NP
authors.lastname: King
authors.affiliation: Department of Chemistry and Biochemistry, UCLA, Los Angeles, California, 90095.|Institute for Genomics and Proteomics, UCLA-DOE, Los Angeles, California, 90095.
authors.forename: Todd O
authors.initials: TO
authors.lastname: Yeates
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington, 98195.|Institute for Protein Design, University of Washington, Seattle, Washington, 98195.|Howard Hughes Medical Institute, University of Washington, Seattle, Washington, 98195.
authors.forename: David
authors.initials: D
authors.lastname: Baker
mesh_terms: D003198:Computer Simulation; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D014644:Genetic Variation; D015202:Protein Engineering; D011506:Proteins / Q000737:chemistry* / Q000235:genetics*; D012995:Solubility
chemical_list: D011506:Proteins
keywords: coassembly; computational protein design; crystal structure; nanomaterial; solubility; symmetry; tetrahedral