Redesigning Protein Cavities as a Strategy for Increasing Affinity in Protein-Protein Interaction: Interferon-γ Receptor 1 as a Model
Joint Authors
Schneider, B.
Černý, Jiří
Biedermannová, Lada
Mikulecký, Pavel
Zahradník, Jiří
Charnavets, Tatsiana
Šebo, Peter
Source
Issue
Vol. 2015, Issue 2015 (31 Dec. 2015), pp.1-12, 12 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2015-04-28
Country of Publication
Egypt
No. of Pages
12
Main Subjects
Abstract EN
Combining computational and experimental tools, we present a new strategy for designing high affinity variants of a binding protein.
The affinity is increased by mutating residues not at the interface, but at positions lining internal cavities of one of the interacting molecules.
Filling the cavities lowers flexibility of the binding protein, possibly reducing entropic penalty of binding.
The approach was tested using the interferon-γ receptor 1 (IFNγR1) complex with IFNγ as a model.
Mutations were selected from 52 amino acid positions lining the IFNγR1 internal cavities by using a protocol based on FoldX prediction of free energy changes.
The final four mutations filling the IFNγR1 cavities and potentially improving the affinity to IFNγ were expressed, purified, and refolded, and their affinity towards IFNγ was measured by SPR.
While individual cavity mutations yielded receptor constructs exhibiting only slight increase of affinity compared to WT, combinations of these mutations with previously characterized variant N96W led to a significant sevenfold increase.
The affinity increase in the high affinity receptor variant N96W+V35L is linked to the restriction of its molecular fluctuations in the unbound state.
The results demonstrate that mutating cavity residues is a viable strategy for designing protein variants with increased affinity.
American Psychological Association (APA)
Černý, Jiří& Biedermannová, Lada& Mikulecký, Pavel& Zahradník, Jiří& Charnavets, Tatsiana& Šebo, Peter…[et al.]. 2015. Redesigning Protein Cavities as a Strategy for Increasing Affinity in Protein-Protein Interaction: Interferon-γ Receptor 1 as a Model. BioMed Research International،Vol. 2015, no. 2015, pp.1-12.
https://search.emarefa.net/detail/BIM-1056481
Modern Language Association (MLA)
Černý, Jiří…[et al.]. Redesigning Protein Cavities as a Strategy for Increasing Affinity in Protein-Protein Interaction: Interferon-γ Receptor 1 as a Model. BioMed Research International No. 2015 (2015), pp.1-12.
https://search.emarefa.net/detail/BIM-1056481
American Medical Association (AMA)
Černý, Jiří& Biedermannová, Lada& Mikulecký, Pavel& Zahradník, Jiří& Charnavets, Tatsiana& Šebo, Peter…[et al.]. Redesigning Protein Cavities as a Strategy for Increasing Affinity in Protein-Protein Interaction: Interferon-γ Receptor 1 as a Model. BioMed Research International. 2015. Vol. 2015, no. 2015, pp.1-12.
https://search.emarefa.net/detail/BIM-1056481
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1056481