[en] We describe a naturally occurring dominant negative P2X1 mutant. This mutant lacks one leucine within a stretch of four leucine residues in its second transmembrane domain (TM2) (amino acids 351-354). Confocal microscopy revealed proper plasma membrane localization of the mutant in stably transfected HEK293 cells. Nevertheless, voltage-clamped HEK293 cells expressing mutated P2X1 channels failed to develop an ATP or ADP-induced current. Furthermore, when co-expressed with the wild type receptor in Xenopus oocytes, the mutated protein exhibited a dose-dependent dominant negative effect on the normal ATP or ADP-induced P2X1 channel activity. These data indicate that deletion of a single apolar amino acid residue at the inner border of the P2X1 TM2 generates a nonfunctional channel.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Oury, Cécile ; Katholieke Universiteit Leuven - KUL > Molecular and Cellular Medicine > CMVB
Toth-Zsamboki, Emese; Katholieke Universiteit Leuven - KUL > Molecular and Cellular Medicine > CMVB
Van Geet, Chris; Katholieke Universiteit Leuven - KUL > Molecular and Cellular Medicine > CMVB
Thijs, Chantal; Katholieke Universiteit Leuven - KUL > Molecular and Cellular Medicine > CMVB
Wei, Li; Katholieke Universiteit Leuven - KUL > Molecular and Cellular Biology > Laboratory of Physiology
Nilius, Bernd; Katholieke Universiteit Leuven - KUL > Molecular and Cellular Medicine > Laboratory of Physiology
Vermylen, Jos; Katholieke Universiteit Leuven - KUL > Molecular and Cellular Medicine > CMVB
Hoylaerts, Marc F; Katholieke Universiteit Leuven - KUL > Molecular and Cellular Biology > CMVB
Language :
English
Title :
A natural dominant negative P2X1 receptor due to deletion of a single amino acid residue.
Publication date :
2000
Journal title :
Journal of Biological Chemistry
ISSN :
0021-9258
eISSN :
1083-351X
Publisher :
American Society for Biochemistry and Molecular Biology, Baltimore, United States - Maryland