Designed asymmetric protein assembly on a symmetric scaffold

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Cellular signaling is regulated by the assembly of proteins into higher-order complexes. Bottom-up creation of synthetic protein assemblies, especially asymmetric complexes, is highly challenging. Presented here is the design and implementation of asymmetric assembly of a ternary protein complex facilitated by Rosetta modeling and thermodynamic analysis. The wild-type symmetric CT32-CT32 interface of the 14-3-3-CT32 complex was targeted, ultimately favoring asymmetric assembly on the 14-3-3 scaffold. Biochemical studies, supported by mass-balance models, allowed characterization of the parameters driving asymmetric assembly. Importantly, our work reveals that both the individual binding affinities and cooperativity between the assembling components are crucial when designing higher-order protein complexes. Enzyme complementation on the 14-3-3 scaffold highlighted that interface engineering of a symmetric ternary complex generates asymmetric protein complexes with new functions.

Original languageEnglish
Pages (from-to)12113-12121
Number of pages9
JournalAngewandte Chemie - International Edition
Volume59
Issue number29
Early online date25 Apr 2020
DOIs
Publication statusPublished - 13 Jul 2020

Keywords

  • cooperativity
  • noncovalent interactions
  • proteins
  • protein–protein interactions
  • self-assembly
  • protein-protein interactions

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