Structure of a designed tetrahedral protein assembly variant engineered to have improved soluble expression

Protein Sci. 2015 Oct;24(10):1695-701. doi: 10.1002/pro.2748. Epub 2015 Aug 6.

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.

Keywords: coassembly; computational protein design; crystal structure; nanomaterial; solubility; symmetry; tetrahedral.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Computer Simulation
  • Crystallography, X-Ray
  • Electrophoresis, Polyacrylamide Gel
  • Gene Expression*
  • Genetic Variation*
  • Protein Engineering*
  • Proteins / chemistry*
  • Proteins / genetics*
  • Solubility

Substances

  • Proteins

Associated data

  • PDB/1nza
  • PDB/1ufy
  • PDB/4ZK7