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

Valentina Elisabetta Viviana Ferrero; Mattia Pedotti; Alessandro Chiadò; Luca Simonelli; Luigi Calzolai; Luca Varani; Teresa Lettieri

doi:10.1371/journal.pone.0102658

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

PLoS One. 2014 Jul 30;9(7):e102658. doi: 10.1371/journal.pone.0102658. eCollection 2014.

Authors

Valentina Elisabetta Viviana Ferrero¹, Mattia Pedotti², Alessandro Chiadò², Luca Simonelli², Luigi Calzolai³, Luca Varani², Teresa Lettieri¹

Affiliations

¹ European Commission - Directorate General - Joint Research Centre, Institute for Environment and Sustainability, Ispra, Varese, Italy.
² Institute for Research in Biomedicine, Bellinzona, Switzerland.
³ European Commission - Directorate General - Joint Research Centre, Institute for Health and Consumer Protection, Ispra, Varese, Italy.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Binding Sites
Ethinyl Estradiol / chemistry
Ethinyl Estradiol / pharmacology
Humans
Molecular Docking Simulation*
Molecular Sequence Data
Point Mutation
Protein Binding
Receptors, Estrogen / chemistry*
Receptors, Estrogen / genetics
Receptors, Estrogen / metabolism

Substances

Receptors, Estrogen
Ethinyl Estradiol

Grants and funding

The whole project achievements were possible thanks to the RADAR FP7 project funds FP7- KBBE/Programme ‘Cooperation’ - Research Theme: ‘Food, agriculture and fisheries, and biotechnology’, project number 265721). AC was supported in part by the Erasmus program. LV acknowledges financial support from SVRI, BSJRP and SNF. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.