Redesigning Protein Cavities as a Strategy for Increasing Affinity in Protein-Protein Interaction: Interferon-gamma Receptor 1 as a Model

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F86652036%3A_____%2F15%3A00447258" target="_blank" >RIV/86652036:_____/15:00447258 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1155/2015/716945" target="_blank" >http://dx.doi.org/10.1155/2015/716945</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1155/2015/716945" target="_blank" >10.1155/2015/716945</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Redesigning Protein Cavities as a Strategy for Increasing Affinity in Protein-Protein Interaction: Interferon-gamma Receptor 1 as a Model

Popis výsledku v původním jazyce

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 ofthe interacting molecules. Filling the cavities lowers flexibility of the binding protein, possibly reducing entropic penalty of binding. The approach was tested using the interferon-gamma receptor 1 (IFN gamma R1) complex with IFN gamma as a model. Mutations were selected from 52 amino acid positions lining the IFN gamma R1 internal cavities by using a protocol based on FoldX prediction of free energy changes. The final four mutations filling the IFN gamma R1 cavities and potentially improving the affinity to IFN gamma were expressed, purified, and refolded, and their affinity towards IFN gamma was measured by SPR. While individual cavity mutations yielded receptor constructs exhibiting only slight increase of affinity compared to WT,

Název v anglickém jazyce

Redesigning Protein Cavities as a Strategy for Increasing Affinity in Protein-Protein Interaction: Interferon-gamma Receptor 1 as a Model

Popis výsledku anglicky

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 ofthe interacting molecules. Filling the cavities lowers flexibility of the binding protein, possibly reducing entropic penalty of binding. The approach was tested using the interferon-gamma receptor 1 (IFN gamma R1) complex with IFN gamma as a model. Mutations were selected from 52 amino acid positions lining the IFN gamma R1 internal cavities by using a protocol based on FoldX prediction of free energy changes. The final four mutations filling the IFN gamma R1 cavities and potentially improving the affinity to IFN gamma were expressed, purified, and refolded, and their affinity towards IFN gamma was measured by SPR. While individual cavity mutations yielded receptor constructs exhibiting only slight increase of affinity compared to WT,

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

EB - Genetika a molekulární biologie

OECD FORD obor

—

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2015

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

BioMed Research International

ISSN

2314-6133

e-ISSN

—

Svazek periodika

716945

Číslo periodika v rámci svazku

28 April 2015

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

12

Strana od-do

—

Kód UT WoS článku

000354293900001

EID výsledku v databázi Scopus

—