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Author Spotlight: A Computational Approach to Decipher Amino Acid Preferences in Multispecific Protein-Protein Interactions
Published on: January 26, 2024
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Promiscuous Protein Binding as a Function of Protein Stability.
Ruth Cohen-Khait1, Orly Dym2, Shelly Hamer-Rogotner2
1Department of Biomolecular Sciences, Weizmann Institute of Science, 76100 Rehovot, Israel.
Structure (London, England : 1993)
|December 7, 2017
Summary
Researchers evolved TEM1 beta-lactamase to bind itself, revealing mutations that alter protein structure and reduce thermal stability. This study uncovers a mechanism for purging unwanted protein interactions.
Area of Science:
- Protein engineering
- Molecular evolution
- Biochemistry
Background:
- Proteins must discriminate between desired and undesired binding partners.
- Understanding the evolution of protein-protein interactions is crucial for protein design and function.
Purpose of the Study:
- To investigate the evolutionary mechanisms governing protein-protein interactions.
- To engineer TEM1 beta-lactamase for self-binding and analyze the resulting structural and stability changes.
Main Methods:
- Yeast surface display was used to select a library of TEM1 beta-lactamase variants.
- X-ray crystallography was employed to determine the structural basis of the evolved interactions.
- Thermal stability assays were performed on engineered mutants.
Main Results:
- Three mutations enabled micromolar affinity binding between TEM1 beta-lactamase variants.
- Selected mutations altered the protein fold, removing an N'-terminal helix and promoting beta-sheet-mediated interactions.
- A key interface mutation (E58V) is analogous to the sickle-cell mutation.
- Engineered mutations reduced the thermal stability of the protein, preventing their insertion into the wild-type.
Conclusions:
- Protein fold modification, rather than direct interface complementarity, can drive self-binding.
- Loss of thermal stability acts as a purging mechanism against undesirable protein interactions.
- Evolutionary constraints can limit the incorporation of beneficial mutations due to stability trade-offs.
Keywords:
TEM1de novo protein interactiondimerizationevolutionmutationspromiscuityprotein-proteinspecificitythermal stabilityβ-sheet augmentationMore Related Videos
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