title: A Residue Quartet in the Extracellular Domain of the Prolactin Receptor Selectively Controls Mitogen-activated Protein Kinase Signaling.
abstract: Cytokine receptors elicit several signaling pathways, but it is poorly understood how they select and discriminate between them. We have scrutinized the prolactin receptor as an archetype model of homodimeric cytokine receptors to address the role of the extracellular membrane proximal domain in signal transfer and pathway selection. Structure-guided manipulation of residues involved in the receptor dimerization interface identified one residue (position 170) that in cell-based assays profoundly altered pathway selectivity and species-specific bio-characteristics. Subsequent in vitro spectroscopic and nuclear magnetic resonance analyses revealed that this residue was part of a residue quartet responsible for specific local structural changes underlying these effects. This included alteration of a novel aromatic T-stack within the membrane proximal domain, which promoted selective signaling affecting primarily the MAPK (ERK1/2) pathway. Importantly, activation of the MAPK pathway correlated with in vitro stabilities of ternary ligand·receptor complexes, suggesting a threshold mean lifetime of the complex necessary to achieve maximal activation. No such dependence was observed for STAT5 signaling. Thus, this study establishes a residue quartet in the extracellular membrane proximal domain of homodimeric cytokine receptors as a key regulator of intracellular signaling discrimination.
authors.affiliation: From the Inserm, U1151, Institut Necker Enfants Malades, Equipe Physiopathologie des Hormones PRL/GH, Paris 75014, France, the Université Paris Descartes, Sorbonne Paris Cité, Paris 75006, France.
authors.forename: Chi
authors.initials: C
authors.lastname: Zhang
authors.affiliation: the Structural Biology and NMR Laboratory, Department of Biology, University of Copenhagen, DK-2200 Copenhagen N, Denmark.
authors.forename: Mads
authors.initials: M
authors.lastname: Nygaard
authors.affiliation: the Structural Biology and NMR Laboratory, Department of Biology, University of Copenhagen, DK-2200 Copenhagen N, Denmark.
authors.forename: Gitte W
authors.initials: GW
authors.lastname: Haxholm
authors.affiliation: From the Inserm, U1151, Institut Necker Enfants Malades, Equipe Physiopathologie des Hormones PRL/GH, Paris 75014, France, the Université Paris Descartes, Sorbonne Paris Cité, Paris 75006, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Boutillon
authors.affiliation: From the Inserm, U1151, Institut Necker Enfants Malades, Equipe Physiopathologie des Hormones PRL/GH, Paris 75014, France, the Université Paris Descartes, Sorbonne Paris Cité, Paris 75006, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Bernadet
authors.affiliation: the Institut Pasteur, Plateforme de Biophysique des Macromolécules et de leurs Interactions, Département de Biologie Structurale et Chimie, Paris 75015, France, and.
authors.forename: Sylviane
authors.initials: S
authors.lastname: Hoos
authors.affiliation: the Institut Pasteur, Plateforme de Biophysique des Macromolécules et de leurs Interactions, Département de Biologie Structurale et Chimie, Paris 75015, France, and.
authors.forename: Patrick
authors.initials: P
authors.lastname: England
authors.affiliation: the Université Paris Descartes, Sorbonne Paris Cité, Paris 75006, France, Laboratoire de Cristallographie et RMN Biologiques CNRS, UMR 8015, Paris 75006, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Broutin
authors.affiliation: the Structural Biology and NMR Laboratory, Department of Biology, University of Copenhagen, DK-2200 Copenhagen N, Denmark.
authors.forename: Birthe B
authors.initials: BB
authors.lastname: Kragelund
authors.affiliation: From the Inserm, U1151, Institut Necker Enfants Malades, Equipe Physiopathologie des Hormones PRL/GH, Paris 75014, France, the Université Paris Descartes, Sorbonne Paris Cité, Paris 75006, France, vincent.goffin@inserm.fr.
authors.forename: Vincent
authors.initials: V
authors.lastname: Goffin
mesh_terms: D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D002942:Circular Dichroism; D016207:Cytokines / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D006801:Humans; D008024:Ligands; D020935:MAP Kinase Signaling System; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011981:Receptors, Prolactin / Q000378:metabolism*; D050799:STAT5 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction; D013050:Spectrometry, Fluorescence
chemical_list: D016207:Cytokines; D008024:Ligands; D011981:Receptors, Prolactin; D050799:STAT5 Transcription Factor; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: Cell Proliferation; Cell Surface Receptor; Cytokine; Extracellular Signal-regulated Kinase (ERK); Nuclear Magnetic Resonance (NMR); STAT Transcription Factor