title: Post-translational allosteric activation of the P2X7 receptor through glycosaminoglycan chains of CD44 proteoglycans.
abstract: Here, we present evidence for the positive allosteric modulation of the P2X7 receptor through glycosaminoglycans (GAGs) in CHO (cell line derived from the ovary of the Chinese hamster) cells. The marked potentiation of P2X7 activity through GAGs in the presence of non-saturating agonists concentrations was evident with the endogenous expression of the receptor in CHO cells. The presence of GAGs on the surface of CHO cells greatly increased the sensitivity to adenosine 5'-triphosphate and changed the main P2X7 receptor kinetic parameters EC50, Hill coefficient and E max. GAGs decreased the allosteric inhibition of P2X7 receptor through Mg(2+). GAGs activated P2X7 receptor-mediated cytoplasmic Ca(2+) influx and pore formation. Consequently, wild-type CHO-K1 cells were 2.5-fold more sensitive to cell death induced through P2X7 agonists than mutant CHO-745 cells defective in GAGs biosynthesis. In the present study, we provide the first evidence that the P2X7 receptor interacts with CD44 on the CHO-K1 cell surface. Thus, these data demonstrated that GAGs positively modulate the P2X7 receptor, and sCD44 is a part of a regulatory positive feedback loop linking P2X7 receptor activation for the intracellular response mediated through P2X7 receptor stimulation.
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo , São Paulo, Brazil.
authors.forename: Gedd
authors.initials: G
authors.lastname: Moura
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo , São Paulo, Brazil.
authors.forename: S V
authors.initials: SV
authors.lastname: Lucena
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo , São Paulo, Brazil.
authors.forename: M A
authors.initials: MA
authors.lastname: Lima
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo, São Paulo, Brazil; Grupo de Pesquisa em Biomateriais e Biotecnologia, Universidade Bandeirante de São Paulo, São Paulo, Brazil.
authors.forename: F D
authors.initials: FD
authors.lastname: Nascimento
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo, São Paulo, Brazil; Division of Developmental Biology, Cincinnati Children's Hospital and Research, Cincinnati, OH, USA.
authors.forename: T F
authors.initials: TF
authors.lastname: Gesteira
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo , São Paulo, Brazil.
authors.forename: H B
authors.initials: HB
authors.lastname: Nader
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo, São Paulo, Brazil; Centro Interdisciplinar de Investigação Bioquıí́mica, Universidade de Mogi das Cruzes, São Paulo, Brazil.
authors.forename: E J
authors.initials: EJ
authors.lastname: Paredes-Gamero
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo, São Paulo, Brazil; Centro Interdisciplinar de Investigação Bioquıí́mica, Universidade de Mogi das Cruzes, São Paulo, Brazil.
authors.forename: Ils
authors.initials: I
authors.lastname: Tersariol