Structure of a designed tetrahedral protein assembly variant engineered to have improved soluble expression.
Publication Type:Journal Article
Source:Protein Sci, Volume 24, Issue 10, p.1695-701 (2015)
Keywords:Computer Simulation, Crystallography, X-Ray, Electrophoresis, Polyacrylamide Gel, Gene Expression, Genetic Variation, Protein Engineering, Proteins, Solubility
<p>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. </p>