Primary structure and expression of a naturally truncated human P2X ATP receptor subunit from brain and immune system

Lê, Khanh-Tuoc; Paquet, Marie‐Eve; Nouel, Dominique; Babinski, K.; Séguéla, Philippe

doi:10.1016/s0014-5793(97)01380-x

Cited by 77 publications

(49 citation statements)

References 28 publications

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“…It has previously been well documented that the form of the receptor that does not contain exon 10 can not form homomeric channels despite apparently good membrane expression (Lê et al, 1997). Human tissues seem to contain more P2X receptor forms that are spliced so as to produce nonfunctional proteins than are found in rodent orthologs (e.g., P2X 2 , Lynch et al, 1999;P2X 6 , Urano et al, 1997;see North, 2002); this might reflect a loss of function during evolution.…”

Section: Discussionmentioning

confidence: 99%

“…The human P2X 5 receptor cDNA was isolated from brain, and its RNA was particularly enriched in thymus and other immune cells (Lê et al, 1997). However, alignment with other members of the P2X families showed that this cDNA lacked exon 10.…”

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confidence: 99%

“…Exon 10 encodes a 22-amino acid segment of protein including much of the second of the two membranespanning domains. This cDNA did not encode functional channels, but a chimeric receptor in which the C-terminal region of the rat receptor (starting at the position equivalent to the beginning of exon 10) was joined to the human receptor provided robust ATP-activated currents when expressed in X. laevis oocytes (Lê et al, 1997).…”

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confidence: 99%

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Pharmacological and Biophysical Properties of the Human P2X₅Receptor

et al. 2003

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We constructed a full-length human P2X 5 purinoceptor cDNA by incorporating a sequence corresponding to exon 10, which is missing in cDNAs cloned previously from human tissues. We studied the functional properties by patch-clamp recording and fluorescence imaging after expression in human embryonic kidney 293 cells. ATP (1-100 M; half-maximal current at 4 M) elicited inward currents at Ϫ60 mV; these persisted during brief (2 s) applications but declined during longer applications. The peak current was dependent on the holding potential and showed little rectification; however, both the desensitization during the application and the decline in the current when ATP was washed out were slower at ϩ30 mV than at Ϫ60 mV. 2Ј,3Ј-O-(4-Benzoyl)-benzoyl-ATP and ␣␤-methylene-ATP mimicked the action of ATP (half-maximal concentrations 6 and 161 M, respectively). The currents were inhibited by suramin, pyridoxal-5-phosphate-6-azo-2Ј,4Ј-disulfonic acid and Brilliant Blue G, with half-maximal inhibition at 3, 0.2, and 0.5 M, respectively; 2Ј,3Ј-O-(2Ј,4Ј,6Ј-trinitrophenol)-ATP (1 M) was ineffective. Removing divalent cations did not significantly alter ATP concentration-response curves. Reversal potential measurements showed that the human P2X 5 receptor was permeable to calcium (P Ca /P Na ϭ 1.5) and N-methyl-D-glucamine (NMDG) (P NMDG /P Na ϭ 0.4); it was also permeable to chloride (P Cl /P Na ϭ 0.5) but not gluconate (P gluc /P Na ϭ 0.01) ions. The permeability to NMDG developed as quickly as the channel opened, in contrast to the P2X 7 receptor where the NMDG permeability develops over several seconds. Cells expressing human P2X 5 receptors also rapidly accumulated the propidium dye YO-PRO-1 in response to ATP.A P2X 5 subunit cDNA was first isolated from libraries of rat sympathetic ganglia (Collo et al., 1996) and heart (Garcia-Guzman et al., 1996). The RNA is expressed predominantly in heart (Garcia-Guzman et al., 1996) but there is also expression in brain, spinal cord, and adrenal gland (GarciaGuzman et al., 1996). Immunoreactivity for P2X 5 subunits has also been described in developing skeletal muscle of the rat (Meyer et al., 1999;Ryten et al., 2001); recently, the receptor has been implicated in the differentiation of satellite cells into mature multinucleated muscle fibers (Ryten et al., 2002). Mammalian skin and endocrine (Glass and Burnstock, 2001) cells also show P2X 5 immunoreactivity.When expressed in HEK293 cells, the rat P2X 5 subunit cDNAs gave rise to currents activated by ATP, but these were very small and their properties not extensively studied. The current was elicited by ATP [half-maximal concentration (EC 50 ) about 10 M] but not by ␣␤meATP, and it was readily blocked by suramin and PPADS in the 1 to 10 M concentration range (Collo et al., 1996;Garcia-Guzman et al., 1996). A mouse cDNA has also been cloned and expressed; again, the currents recorded from HEK293 cells were only a few tens of picoamperes and they were not studied in detail . On the other hand, although these two mammalian recep...

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Section: Discussionmentioning

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Pharmacological and Biophysical Properties of the Human P2X₅Receptor

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“…In humans, the expression and function of this receptor is still unclear. The mRNA transcripts have been detected predominantly in tissues related to the immune system (Lê et al, 1997). The recombinant rat and zP2X5R generates a low-amplitude nondesensitizing current, whereas P2X5Rs from other species respond to ATP with large, rapidly activating and slowly desensitizing currents.…”

Section: Activation and Regulation Of P2xrsmentioning

confidence: 99%

Activation and Regulation of Purinergic P2X Receptor Channels

et al. 2011

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“…One interesting example is the disappearance of Ij intron of P2X5 receptor in primates (Mmu, Ggg, Cja, Hsa, Ptr). Lost of an exon (and adjacent Ij intron) of P2X5 in primates, which partially constitutes TM2 domain, usually leads to nonfunctional proteins [1,54,55].…”

Section: Phylogenetic Analysis Of the Putative P2x Genesmentioning

confidence: 99%

Comparative study of the P2X gene family in animals and plants

Hou

Cao

2016

Purinergic Signalling

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P2X receptors are ligand-gated ion channels that can bind with the adenosine triphosphate (ATP) and have diverse functional roles in neuropathic pain, inflammation, special sense, and so on. In this study, 180 putative P2X genes, including 176 members in 32 animal species and 4 members in 3 species of lower plants, were identified. These genes were divided into 13 groups, including 7 groups in vertebrates and 6 groups in invertebrates and lower plants, through phylogenetic analysis. Their gene organization and motif composition are conserved in most predicted P2X members, while groupspecific features were also found. Moreover, synteny relationships of the putative P2X genes in vertebrates are conserved while simultaneously experiencing a series of gene insertion, inversion, and transposition. Recombination signals were detected in almost all of the vertebrates and invertebrates, suggesting that intragenic recombination may play a significant role in the evolution of P2X genes. Selection analysis also identified some positively selected sites that acted on the evolution of most of the predicted P2X proteins. The phenomenon of alternative splicing occurred commonly in the putative P2X genes of vertebrates. This article explored in depth the evolutional relationship among different subtypes of P2X genes in animal and plants and might serve as a solid foundation for deciphering their functions in further studies.

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Primary structure and expression of a naturally truncated human P2X ATP receptor subunit from brain and immune system

Cited by 77 publications

References 28 publications

Pharmacological and Biophysical Properties of the Human P2X₅Receptor

Pharmacological and Biophysical Properties of the Human P2X₅Receptor

Activation and Regulation of Purinergic P2X Receptor Channels

Comparative study of the P2X gene family in animals and plants

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Primary structure and expression of a naturally truncated human P2X ATP receptor subunit from brain and immune system

Cited by 77 publications

References 28 publications

Pharmacological and Biophysical Properties of the Human P2X5Receptor

Pharmacological and Biophysical Properties of the Human P2X5Receptor

Activation and Regulation of Purinergic P2X Receptor Channels

Comparative study of the P2X gene family in animals and plants

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Product

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Pharmacological and Biophysical Properties of the Human P2X₅Receptor

Pharmacological and Biophysical Properties of the Human P2X₅Receptor