The intracellular tyrosine residues of the ATP‐gated P2X1 ion channel are essential for its function

FEBS Letters

Oury, Cecile¹  Vermylen, Jos¹  Watanabe, Hiroyuki²  Hoylaerts, Marc F¹  Nilius, Bernd²  Toth-Zsamboki, Emese¹ 

[1] Center for Molecular and Vascular Biology, University of Leuven, Herestraat 49, 3000 Leuven, Belgium;Laboratory of Physiology, University of Leuven, Herestraat 49, 3000 Leuven, Belgium

关键词: P2X1 receptor;    ATP-gated non-selective cation channel;    Tyrosine residue;    Site-directed mutagenesis;    Three-dimensional structure;   

DOI  :  10.1016/S0014-5793(02)02987-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The four highly conserved intracellular tyrosine residues of the P2X₁ ion channel were mutated into phenylalanine. Simultaneous electrophysiological and calcium measurements in transfected human embryonic kidney (HEK 293) cells indicated that Y362F and Y370F mutants were non-functional, despite their proper plasma membrane expression. The Y16F and Y363F mutants retained 2.2% and 26% of the wild-type P2X₁ activity, respectively. However, no tyrosine phosphorylation was detected on Western blots of P2X₁ immunoprecipitates derived either from HEK 293 cell lysates or from human platelets, expressing P2X₁ endogenously. Thus, Y16, Y362, Y363 and Y370 are required for the appropriate three-dimensional structure and function of the intracellular P2X₁ domains.

【 授权许可】

Unknown