Increasing Affinity of Interferon-γ Receptor 1 to Interferon-γ by Computer-Aided Design
Joint Authors
Schneider, B.
Mikulecký, Pavel
Černý, Jiří
Biedermannová, Lada
Petroková, Hana
Kuchař, Milan
Vondrášek, Jiří
Malý, Petr
Šebo, Peter
Source
Issue
Vol. 2013, Issue 2013 (31 Dec. 2013), pp.1-12, 12 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2013-10-02
Country of Publication
Egypt
No. of Pages
12
Main Subjects
Abstract EN
We describe a computer-based protocol to design protein mutations increasing binding affinity between ligand and its receptor.
The method was applied to mutate interferon-γ receptor 1 (IFN-γ-Rx) to increase its affinity to natural ligand IFN-γ, protein important for innate immunity.
We analyzed all four available crystal structures of the IFN-γ-Rx/IFN-γ complex to identify 40 receptor residues forming the interface with IFN-γ.
For these 40 residues, we performed computational mutation analysis by substituting each of the interface receptor residues by the remaining standard amino acids.
The corresponding changes of the free energy were calculated by a protocol consisting of FoldX and molecular dynamics calculations.
Based on the computed changes of the free energy and on sequence conservation criteria obtained by the analysis of 32 receptor sequences from 19 different species, we selected 14 receptor variants predicted to increase the receptor affinity to IFN-γ.
These variants were expressed as recombinant proteins in Escherichia coli, and their affinities to IFN-γ were determined experimentally by surface plasmon resonance (SPR).
The SPR measurements showed that the simple computational protocol succeeded in finding two receptor variants with affinity to IFN-γ increased about fivefold compared to the wild-type receptor.
American Psychological Association (APA)
Mikulecký, Pavel& Černý, Jiří& Biedermannová, Lada& Petroková, Hana& Kuchař, Milan& Vondrášek, Jiří…[et al.]. 2013. Increasing Affinity of Interferon-γ Receptor 1 to Interferon-γ by Computer-Aided Design. BioMed Research International،Vol. 2013, no. 2013, pp.1-12.
https://search.emarefa.net/detail/BIM-1005034
Modern Language Association (MLA)
Mikulecký, Pavel…[et al.]. Increasing Affinity of Interferon-γ Receptor 1 to Interferon-γ by Computer-Aided Design. BioMed Research International No. 2013 (2013), pp.1-12.
https://search.emarefa.net/detail/BIM-1005034
American Medical Association (AMA)
Mikulecký, Pavel& Černý, Jiří& Biedermannová, Lada& Petroková, Hana& Kuchař, Milan& Vondrášek, Jiří…[et al.]. Increasing Affinity of Interferon-γ Receptor 1 to Interferon-γ by Computer-Aided Design. BioMed Research International. 2013. Vol. 2013, no. 2013, pp.1-12.
https://search.emarefa.net/detail/BIM-1005034
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1005034