Structural characterization of the stem-stem dimerization interface between prolactin receptor chains complexed with the natural hormone

Autor: Vincent Goffin, Johannes F. Van Agthoven, Chi Zhang, Isabelle Broutin, Bruno Baron, Bertrand Raynal, Patrick England, Estelle Tallet, Sylviane Hoos
Přispěvatelé: Laboratoire de cristallographie et RMN biologiques (LCRB - UMR 8015), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre de recherche Croissance et signalisation (UMR_S 845), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5), Biophysique des Macromolécules et de leurs Interactions, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), This work was supported, in part, by University Paris Descartes and the Agence Nationale de la Recherche (grant ANR-07-PCVI-0029). J.v.A., C.Z., and E.T. acknowledge the Ministère de l'Education Nationale de la Recherche et de la Technologie for fellowship support., ANR-07-PCVI-0029,PROLACTUMOR,Processus physiopathologiques impliquant le récepteur de la prolactine : approches physico-chimiques dynamiques et structurales(2007), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Rok vydání: 2010
Předmět:
Models
Molecular
MESH: Protein Structure
Quaternary
Receptors
Prolactin
[SDV]Life Sciences [q-bio]
Protein Data Bank (RCSB PDB)
MESH: Prolactin
Growth hormone receptor
Biology
Crystallography
X-Ray
03 medical and health sciences
Paracrine signalling
MESH: Receptors
Prolactin
0302 clinical medicine
Structural Biology
MESH: Protein Binding
Receptor
Autocrine signalling
Protein Structure
Quaternary
Molecular Biology
030304 developmental biology
0303 health sciences
MESH: Protein Multimerization
Prolactin receptor
Mutagenesis
Surface Plasmon Resonance
MESH: Crystallography
X-Ray
MESH: Amino Acid Substitution
MESH: Surface Plasmon Resonance
Cell biology
Prolactin
MESH: Mutagenesis
Site-Directed
Biochemistry
Amino Acid Substitution
030220 oncology & carcinogenesis
Growth hormone receptor complex
Mutagenesis
Site-Directed
Protein Multimerization
MESH: Models
Molecular
Protein Binding
Zdroj: Journal of Molecular Biology
Journal of Molecular Biology, Elsevier, 2010, 404 (1), pp.112-126. ⟨10.1016/j.jmb.2010.09.036⟩
Journal of Molecular Biology, 2010, 404 (1), pp.112-126. ⟨10.1016/j.jmb.2010.09.036⟩
ISSN: 1089-8638
0022-2836
DOI: 10.1016/j.jmb.2010.09.036⟩
Popis: International audience; The most promising approach to targeting the tumor-growth-promoting actions of prolactin (PRL) mediated by its autocrine/paracrine pathway has been the development of specific PRL receptor (PRLR) antagonists. However, the optimization of such antagonists requires a thorough understanding of the activation mechanism of PRLR. We have thus conducted a systematic X-ray crystallographic study in order to visualize the successive steps of PRLR activation by PRL. We report here the structure at 3.35 Å resolution of the 1:2 complex between natural PRL and two PRLR chains (PRLR1 and PRLR2), corresponding to the final activated state of PRLR. Further than our previously published structure involving an affinity-matured PRL variant, this structure allowed to visualize for the first time the loop L5 spanning PRLR2 residues Thr133-Phe140, revealing its central implication for the three intermolecular interfaces of the complex. We equally succeeded in obtaining a comprehensive picture of the PRLR-PRLR dimerization interface, also called stem-stem interface. Site-directed mutagenesis was conducted to probe the energetic importance of stem-stem contacts highlighted by the structure. Surprisingly, in spite of significant structural differences between the PRL/PRLR(2) complex and the 1:2 growth hormone/growth hormone receptor complex, our mutational data suggest that hot-spot residues that stabilize the receptor dimerization interface are equivalent in the two complexes. This study provides a new overall picture of the structural features of PRLR involved in stabilizing its complex with PRL.
Databáze: OpenAIRE
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