title: Identification of transmembrane domains that regulate spatial arrangements and activity of prokineticin receptor 2 dimers.
abstract: The chemokine prokineticin 2 (PK2) activates its cognate G protein-coupled receptor (GPCR) PKR2 to elicit various downstream signaling pathways involved in diverse biological processes. Many GPCRs undergo dimerization that can modulate a number of functions including membrane delivery and signal transduction. The aim of this study was to elucidate the interface of PKR2 protomers within dimers by analyzing the ability of PKR2 transmembrane (TM) deletion mutants to associate with wild type (WT) PKR2 in yeast using co-immunoprecipitation and mammalian cells using bioluminescence resonance energy transfer. Deletion of TMs 5-7 resulted in a lack of detectable association with WT PKR2, but could associate with a truncated mutant lacking TMs 6-7 (TM1-5). Interestingly, TM1-5 modulated the distance, or organization, between protomers and positively regulated Gαs signaling and surface expression of WT PKR2. We propose that PKR2 protomers form type II dimers involving TMs 4 and 5, with a role for TM5 in modulation of PKR2 function.
authors.affiliation: Department of Biochemical Science, Sapienza Università di Roma, Italy; Department of Surgery and Cancer, Institute of Reproductive and Developmental Biology, Imperial College London, UK.
authors.forename: S
authors.initials: S
authors.lastname: Sposini
authors.affiliation: Laboratori de Medicina Computacional, Unitat de Bioestadística, Facultat de Medicina, Universitat Autònoma de Barcelona, Spain.
authors.forename: G
authors.initials: G
authors.lastname: Caltabiano
authors.affiliation: Department of Surgery and Cancer, Institute of Reproductive and Developmental Biology, Imperial College London, UK. Electronic address: a.hanyaloglu@imperial.ac.uk.
authors.forename: A C
authors.initials: AC
authors.lastname: Hanyaloglu
authors.affiliation: Department of Biochemical Science, Sapienza Università di Roma, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Miele
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D057809:HEK293 Cells; D006801:Humans; D055503:Protein Multimerization / Q000502:physiology*; D017434:Protein Structure, Tertiary; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D018000:Receptors, Peptide / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion
chemical_list: C470838:PROKR2 protein, human; D043562:Receptors, G-Protein-Coupled; D018000:Receptors, Peptide
keywords: Bioluminescence resonance energy transfer (BRET); Dimerization; G-protein coupled receptor (GPCR); Molecular modeling; Prokineticin receptor 2 (PKR2); Signaling