title: Rational modification of estrogen receptor by combination of computational and experimental analysis.
abstract: In this manuscript, we modulate the binding properties of estrogen receptor protein by rationally modifying the amino acid composition of its ligand binding domain. By combining sequence alignment and structural analysis of known estrogen receptor-ligand complexes with computational analysis, we were able to predict estrogen receptor mutants with altered binding properties. These predictions were experimentally confirmed by producing single point variants with up to an order of magnitude increased binding affinity towards some estrogen disrupting chemicals and reaching an half maximal inhibitory concentration (IC50) value of 2 nM for the 17α-ethinylestradiol ligand. Due to increased affinity and stability, utilizing such mutated estrogen receptor instead of the wild type as bio-recognition element would be beneficial in an assay or biosensor.
authors: European Commission - Directorate General - Joint Research Centre, Institute for Environment and Sustainability, Ispra, Varese, Italy.
authors: Valentina Elisabetta Viviana
authors: VE
authors: Ferrero
authors: Institute for Research in Biomedicine, Bellinzona, Switzerland.
authors: Mattia
authors: M
authors: Pedotti
authors: Institute for Research in Biomedicine, Bellinzona, Switzerland.
authors: Alessandro
authors: A
authors: Chiadò
authors: Institute for Research in Biomedicine, Bellinzona, Switzerland.
authors: Luca
authors: L
authors: Simonelli
authors: European Commission - Directorate General - Joint Research Centre, Institute for Health and Consumer Protection, Ispra, Varese, Italy.
authors: Luigi
authors: L
authors: Calzolai
authors: Institute for Research in Biomedicine, Bellinzona, Switzerland.
authors: Luca
authors: L
authors: Varani
authors: European Commission - Directorate General - Joint Research Centre, Institute for Environment and Sustainability, Ispra, Varese, Italy.
authors: Teresa
authors: T
authors: Lettieri
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D004997:Ethinyl Estradiol / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D017354:Point Mutation; D011485:Protein Binding; D011960:Receptors, Estrogen / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D011960:Receptors, Estrogen; D004997:Ethinyl Estradiol