Identification of P2X2/P2X4/P2X6 heterotrimeric receptors using atomic force microscopy (AFM) imaging

Autor(es)

Antonio, Ligia S. ^{[1, 2]} ; Stewart, Andrew P. ^[1] ; Varanda, Wamberto A. ^[2] ; Edwardson, J. Michael ^[1]

Afiliação do(s) autor(es)

^[1] Univ Cambridge, Dept Pharmacol, Cambridge CB2 1PD - England

^[2] Univ Sao Paulo, Dept Physiol, Sch Med Ribeirao Preto, BR-14049 Ribeirao Preto, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: FEBS Letters; v. 588, n. 12, p. 2125-2128, JUN 5 2014.
• Citações Web of Science: 7

Resumo

Seven P2X purinergic receptor subunits have been identified: P2X1-P2X7. The overlapping expression of P2X2, P2X4 and P2X6 subunits has been shown in different cell types, and functional analysis of P2X receptors in Leydig cells suggests that the three subunits might interact. Here, His(6)-tagged P2X2, HA-tagged P2X4 and FLAG-tagged P2X6 subunits were co-expressed in tsA 201 cells. After sequential co-immunoprecipitation using anti-HA and anti-FLAG beads, all three subunits were present, demonstrating their interaction. Atomic force microscopy (AFM) imaging revealed receptors that were specifically decorated by both an anti-His(6) antibody and an anti-HA Fab fragment, indicating the presence of a P2X2/4/6 heterotrimer. To our knowledge, this is the first report of a P2X receptor containing three different subunits. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 08/10000-0 - Expressão de subunidades P2X em células de Leydig de camundongos em diferentes idades e interações entre as subunidades P2X2, P2X4, P2X6 formadoras de receptores funcionais
• Beneficiário: Ligia Subitoni Antonio
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 06/50954-7 - Correntes iônicas e receptores na fisiologia de células excitáveis e não excitáveis
• Beneficiário: Wamberto Antonio Varanda
• Modalidade de apoio: Auxílio à Pesquisa - Temático