title: Production of functional recombinant G-protein coupled receptors for heteromerization studies.
abstract: G-protein-coupled receptors (GPCRs) represent a diverse protein family of receptors that transduce signals from the extracellular surrounding to intracellular signaling molecules evoking various cellular responses. It is now widely accepted that GPCRs are expressed and function as dimers or most probably as oligomers of more than two receptor protomers. The heteromer has different biochemical and pharmacological characteristics from the monomers, which increases the functional responses of GPCRs. GPCRs are involved in many diseases, and are also the target of around half of all modern medicinal drugs. In the case of Parkinson's disease, a degenerative process caused by gradual disappearance of dopaminergic nigrostriatal neurons, it is suspected that the targets for treatment should be dopamine-receptor-containing heteromers. Technologies based on the use of fluorescent- or luminescent-fused receptors and adaptations of resonance energy transfer (RET) techniques have been useful in investigating the functional inter-relationships between receptors in a heteromer. In this study functional recombinant adenosine A(2A)-Rluc, dopamine D(2)-GFP(2) and histamine H(3)-YFP receptor fusion proteins were successfully cloned and characterized, producing the essential basis for heteromerization studies between these receptors. This might provide a better insight into their pharmacological and functional inter-relationships in the brain and enable the design and evaluation of new therapeutic strategies for Parkinson's disease.
authors.affiliation: Department of Experimental Oncology, Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia. milena.cavic@ncrc.ac.rs
authors.forename: Milena
authors.initials: M
authors.lastname: Cavić
authors.forename: Carme
authors.initials: C
authors.lastname: Lluís
authors.forename: Estefanía
authors.initials: E
authors.lastname: Moreno
authors.forename: Jana
authors.initials: J
authors.lastname: Bakešová
authors.forename: Enric I
authors.initials: EI
authors.lastname: Canela
authors.forename: Gemma
authors.initials: G
authors.lastname: Navarro
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D018491:Dopamine Agonists / Q000494:pharmacology; D015195:Drug Design; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D010300:Parkinson Disease / Q000188:drug therapy / Q000378:metabolism; D055503:Protein Multimerization / Q000235:genetics*; D043705:Receptor, Adenosine A2A / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D017448:Receptors, Dopamine D2 / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D018100:Receptors, Histamine H3 / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D018491:Dopamine Agonists; D008164:Luminescent Proteins; D043705:Receptor, Adenosine A2A; D017448:Receptors, Dopamine D2; D043562:Receptors, G-Protein-Coupled; D018100:Receptors, Histamine H3; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins