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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 17?-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.

SUBMITTER: Ferrero VE

PROVIDER: S-EPMC4116177 | biostudies-other | 2014

REPOSITORIES: biostudies-other

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Rational modification of estrogen receptor by combination of computational and experimental analysis.

Ferrero Valentina Elisabetta Viviana VE Pedotti Mattia M Chiadò Alessandro A Simonelli Luca L Calzolai Luigi L Varani Luca L Lettieri Teresa T

PloS one 20140730 7

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 increas ...[more]

PMID: 25075862

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