Mutation of putative N-linked glycosylation sites on the human nucleotide receptor P2X7 reveals a key residue important for receptor function.
Mutation of putative N-linked glycosylation sites on the human nucleotide receptor P2X7 reveals a key residue important for receptor function.
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DOI：
10.1021/bi902083n
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发表时间：
2010-06-08
期刊：
影响因子：
2.9
通讯作者：
Bertics, Paul J.
中科院分区：
文献类型：
--
作者：
Lenertz, Lisa Y.;Wang, Ziyi;Guadarrama, Arturo;Hill, Lindsay M.;Gavala, Monica L.;Bertics, Paul J.
The nucleotide receptor P2X7 is an immunomodulatory cation channel and a potential therapeutic target. P2X7 is expressed in immune cells such as monocytes/macrophages and is activated by extracellular ATP following tissue injury or infection. Ligand binding to P2X7 can stimulate ERK1/2, the transcription factor CREB, enzymes linked to the production of reactive oxygen species and interleukin-1 isoforms, and the formation of a non-specific pore. However, little is known about the biochemistry of P2X7, including whether the receptor is N-linked glycosylated and if this modification affects receptor function. Here we provide evidence that P2X7 is sensitive to the glycosidases EndoH and PNGase F, and that the human receptor appears glycosylated on N187, N202, N213, N241 and N284. Mutation of N187 results in diminished P2X7 agonist-induced phosphorylation of ERK1/2, CREB, and p90 ribosomal S6 kinase, as well as decreased pore formation. In further support of a role for glycosylation in receptor function, treatment of RAW 264.7 macrophages with the N-linked glycosylation synthesis inhibitor tunicamycin attenuates P2X7 agonist-induced, but not phorbol ester-induced, ERK1/2 phosphorylation. Interestingly, residue N187 belongs to an N-linked glycosylation consensus sequence found in six of the seven P2X family members, suggesting this site is fundamentally important to P2X receptor function. To address the mechanism whereby N187 mutation attenuates receptor activity, we developed a live cell proteinase K digestion assay that demonstrated altered cell surface expression of P2X7 N187A. This is the first report to map human P2X7 glycosylation sites and reveal residue N187 is critical for receptor trafficking and function.
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影响因子：
5.5
作者：
Gavala, Monica L.;Pfeiffer, Zachary A.;Bertics, Paul J.
通讯作者：
Bertics, Paul J.
影响因子：
5.5
作者：
Humphreys, BD;Dubyak, GR
通讯作者：
Dubyak, GR
DOI：
10.1152/ajpcell.2000.279.4.c1189
发表时间：
2000-10-01
影响因子：
5.5
作者：
Gu, BJ;Zhang, WY;Wiley, JS
通讯作者：
Wiley, JS
影响因子：
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作者：
Dell'Antonio, G;Quattrini, A;Ferrero, ME
通讯作者：
Ferrero, ME
影响因子：
3.5
作者：
Myrtek, Daniel;Idzko, Marco
通讯作者：
Idzko, Marco