title: Application of Circular Dichroism Spectroscopy to the Analysis of the Interaction Between the Estrogen Receptor Alpha and Coactivators: The Case of Calmodulin.
abstract: The estrogen receptor α ligand-binding domain (ERα-LBD) binds the natural hormone 17β-estradiol (E2) to induce transcription and cell proliferation. This process occurs with the contribution of protein and peptide partners (also called coactivators) that can modulate the structure of ERα, and therefore its specificity of action. As with most transcription factors, ERα exhibits a high content of α helix, making it difficult to routinely run spectroscopic studies capable of deciphering the secondary structure of the different partners under binding conditions. Ca(2+)-calmodulin, a protein also highly structured in α-helix, is a key coactivator for ERα activity. Here, we show how circular dichroism can be used to study the interaction of ERα with Ca(2+)-calmodulin. Our approach allows the determination not only of the conformational changes induced upon complex formation but also the dissociation constant (K d) of this interaction.
authors.affiliation: Laboratoire des Biomolécules, Sorbonne Universités, UPMC Univ Paris 06, CNRS-UMR 7203, Université Pierre et Marie Curie-Ecole Normale Supérieure, Tour 23-33, 4, Place Jussieu, 75252, Paris Cedex 05, France.
authors.forename: Emeric
authors.initials: E
authors.lastname: Miclet
authors.affiliation: Plateforme Protéomique PROMETHEE, IAB, University Grenoble Alpes-INSERM, Institut de Biologie et de Pathologie,Grenoble, France; Equipe deSpectrométrie de masse, Institut Parisien de Chimie Moléculaire, UMR 7201, UPMC Univ Paris 06, Paris, France.|INSERM, IAB, Plateforme Protéomique PROMETHEE, 38000, Grenoble, France.|CHU de Grenoble, IAB, Institut de Biologie et de Pathologie, Plateforme Protéomique PROMETHEE, 38000, Grenoble, France.|Equipe de Spectrométrie de masse, Institut Parisien de Chimie Moléculaire, UMR 7201, UPMC University Paris 06, 4, Place Jussieu, 75252, Paris Cedex 05, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Bourgoin-Voillard
authors.affiliation: Laboratoire des Biomolécules, Sorbonne Universités, UPMC Univ Paris 06, CNRS-UMR 7203, Université Pierre et Marie Curie-Ecole Normale Supérieure, Tour 23-33, 4, Place Jussieu, 75252, Paris Cedex 05, France.
authors.forename: Cillian
authors.initials: C
authors.lastname: Byrne
authors.affiliation: Laboratoire des Biomolécules, Sorbonne Universités, UPMC Univ Paris 06, CNRS-UMR 7203, Université Pierre et Marie Curie-Ecole Normale Supérieure, Tour 23-33, 4, Place Jussieu, 75252, Paris Cedex 05, France. yves.jacquot@upmc.fr.
authors.forename: Yves
authors.initials: Y
authors.lastname: Jacquot
mesh_terms: D002147:Calmodulin / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D047628:Estrogen Receptor alpha / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D021241:Spectrometry, Mass, Electrospray Ionization; D013329:Structure-Activity Relationship; D057188:Workflow
chemical_list: D002147:Calmodulin; C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha
keywords: Calmodulin; Circular dichroism; Estrogen receptor; Helix content; K d; Peptide