Increasing Affinity of Interferon-[gamma] Receptor 1 to Interferon-[gamma] by Computer-Aided Design.
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-¿ receptor 1 (IFN-¿-Rx) to increase its affinity to natural ligand IFN-¿, protein important for innate immunity. We analyzed all...
| Publicado en: | BioMed Research International Vol. 2013; pp. 752514 - 752515 |
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| Autores principales: | , , , , , , , , |
| Formato: | Journal Article |
| Publicado: |
Wiley-Blackwell
2013
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=104111759&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 104111759 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 23146133 FT2T jtl: BioMed Research International issn: 23146133 maglogo: N pubinfo: dt: 2013 vid: 2013 pid: 480 pub: Wiley-Blackwell place: Malden, Massachusetts artinfo: ui: 104111759 2012362709 NLM24199198 PMC3807708 104111759 ppf: 752514 ppct: 1 formats: fmt: @attributes: type: P tig: atl: Increasing Affinity of Interferon-[gamma] Receptor 1 to Interferon-[gamma] by Computer-Aided Design. aug: au: Mikulecky, Pavel Cerny, Jií Biedermannová, Lada Petroková, Hana Kucha, Milan Vondrásek, Jií Maly, Petr Sebo, Peter Schneider, Bohdan affil: Institute of Biotechnology AS CR, v. v. i., Vídeská 1083, 142 20 Prague, Czech Republic. sug: subj: Interferons Computer Simulation Biochemical Phenomena Receptors, Cell Surface Amino Acids Surface Plasmon Resonance ab: 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-¿ receptor 1 (IFN-¿-Rx) to increase its affinity to natural ligand IFN-¿, protein important for innate immunity. We analyzed all four available crystal structures of the IFN-¿-Rx/IFN-¿ complex to identify 40 receptor residues forming the interface with IFN-¿. 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-¿. These variants were expressed as recombinant proteins in Escherichia coli, and their affinities to IFN-¿ 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-¿ increased about fivefold compared to the wild-type receptor. pubtype: Academic Journal doctype: Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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