title: Functional and molecular evidence for heteromeric association of P2Y1 receptor with P2Y2 and P2Y4 receptors in mouse granulocytes.
abstract: All hematopoietic cells express P2 receptors, however pharmacological characteristics such as expression and affinity in granulocytes are unknown.
  
  
  
  Pharmacological characteristics of P2 receptors were evaluated by Ca(2+) measurements using Fura-2 fluorophore. P2 receptors expression were analyzed by flow cytometry and RT-PCR. P2 interaction were shown by coimmunoprecipitation, western blotting and FRET.
  
  
  
  Granulocytes were responsive to P2Y agonists, whereas P2X agonists were ineffective. Ca(2+) increase, elicited by ADP and UTP was dependent on intracellular stocks and sensitive to G-coupled receptor inhibition. Moreover, MRS2179, a specific antagonist of the P2Y1 receptor, abolished ADP response. Interestingly, ADP and UTP exhibited full heterologous desensitization, suggesting that these agonists interact with the same receptor. The heteromeric association between P2Y1 receptor and the P2Y2 and P2Y4 receptors was shown by immunoprecipitation and FRET analysis.
  
  
  
  Clear evidence of heteromeric association of P2Y receptors was found during the evaluation of P2 receptors present in mice granulocytes, which could impact in the classical pharmacology of P2Y receptors in granulocytes.
authors.affiliation: Centro Interdisciplinar de Investigação Bioquı́mica, Universidade de Mogi das Cruzes, Av. Dr Cândido Xavier de Almeida Souza, 200, Mogi das Cruzes, São Paulo, Brazil.
authors.forename: Antonio Carlos
authors.initials: AC
authors.lastname: Ribeiro-Filho
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo, Rua Pedro de Toledo, 669 - 9° andar - Prédio de Pesquisa II, R. Três de Maio 100, São Paulo, São Paulo, Brazil.
authors.forename: Marcus Vinicius
authors.initials: MV
authors.lastname: Buri
authors.affiliation: Departamento de Nutrição, Universidade Federal de Pelotas, R. Gomes Carneiro, n°1, 96010-610, Pelotas, Rio Grande do Sul, Brazil.
authors.forename: Carlos Castilho
authors.initials: CC
authors.lastname: Barros
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo, Rua Pedro de Toledo, 669 - 9° andar - Prédio de Pesquisa II, R. Três de Maio 100, São Paulo, São Paulo, Brazil.
authors.forename: Juliana Luporini
authors.initials: JL
authors.lastname: Dreyfuss
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo, Rua Pedro de Toledo, 669 - 9° andar - Prédio de Pesquisa II, R. Três de Maio 100, São Paulo, São Paulo, Brazil.
authors.forename: Helena Bonciani
authors.initials: HB
authors.lastname: Nader
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo, Rua Pedro de Toledo, 669 - 9° andar - Prédio de Pesquisa II, R. Três de Maio 100, São Paulo, São Paulo, Brazil.
authors.forename: Giselle Zenker
authors.initials: GZ
authors.lastname: Justo
authors.affiliation: Departamento de Biofísica, Universidade Federal de São Paulo, R. Botucatu 862, São Paulo, São Paulo, Brazil.
authors.forename: Rogério Bastos
authors.initials: RB
authors.lastname: Craveiro
authors.affiliation: Departamento de Biofísica, Universidade Federal de São Paulo, R. Botucatu 862, São Paulo, São Paulo, Brazil.
authors.forename: João Bosco
authors.initials: JB
authors.lastname: Pesquero
authors.affiliation: Departamento de Biofísica, Universidade Federal de São Paulo, R. Botucatu 862, São Paulo, São Paulo, Brazil.
authors.forename: Antonio
authors.initials: A
authors.lastname: Miranda
authors.affiliation: Departamento de Biofísica, Universidade Federal de São Paulo, R. Botucatu 862, São Paulo, São Paulo, Brazil.
authors.forename: Alice Teixeira
authors.initials: AT
authors.lastname: Ferreira
authors.affiliation: Centro Interdisciplinar de Investigação Bioquı́mica, Universidade de Mogi das Cruzes, Av. Dr Cândido Xavier de Almeida Souza, 200, Mogi das Cruzes, São Paulo, Brazil. edgar.gamero@unifesp.br.|Departamento de Bioquímica, Universidade Federal de São Paulo, Rua Pedro de Toledo, 669 - 9° andar - Prédio de Pesquisa II, R. Três de Maio 100, São Paulo, São Paulo, Brazil. edgar.gamero@unifesp.br.
authors.forename: Edgar Julian
authors.initials: EJ
authors.lastname: Paredes-Gamero
mesh_terms: D000818:Animals; D005260:Female; D005434:Flow Cytometry; D006098:Granulocytes / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D011485:Protein Binding / Q000502:physiology; D058913:Purinergic Agonists / Q000494:pharmacology; D018048:Receptors, Purinergic P2 / Q000737:chemistry / Q000502:physiology*; D058472:Receptors, Purinergic P2Y1 / Q000737:chemistry / Q000502:physiology*; D058473:Receptors, Purinergic P2Y2 / Q000737:chemistry / Q000502:physiology*; D013234:Stem Cells / Q000187:drug effects / Q000502:physiology
chemical_list: D058913:Purinergic Agonists; D018048:Receptors, Purinergic P2; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; C104749:purinoceptor P2Y4
keywords: Calcium signaling; Granulocytes; Heteromeric association; P2Y receptors