TY  - JOUR
AU  - Rokić, Miloš
AU  - Tvrdonova, V.
AU  - Vavra, V.
AU  - Jindrichova, M.
AU  - Obsil, T.
AU  - Stojilković, S. S.
AU  - Zemkova, H.
PY  - 2010
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/403
AB  - Mammalian P2X receptors contain 10 conserved cysteine residues in their ectodomains, which form five disulfide bonds (SS1-5). Here, we analyzed the relevance of these disulfide pairs in rat P2X4 receptor function by replacing one or both cysteines with alanine or threonine, expressing receptors in HEK293 cells and studying their responsiveness to ATP in the absence and presence of ivermectin, an allostenic modulator of these channels. Response to ATP was not altered when both cysteines forming the SS3 bond (C132-C159) were replaced with threonines. Replacement of SS1 (C116-C165), SS2 (C126-C149) and SS4 (C217-C227), but not SS5 (C261-C270), cysteine pairs with threonines resulted in decreased sensitivity to ATP and faster deactivation times. The maximum current amplitude was reduced in SS2, SS4 and SS5 double mutants and could be partially rescued by ivermectin in SS2 and SS5 double mutants. This response pattern was also observed in numerous single residue mutants, but receptor function was not affected when the 217 cysteine was replaced with threonine or arginine or when the 261 cysteine was replaced with alanine. These results suggest that the SS1, SS2 and SS4 bonds contribute substantially to the structure of the ligand binding pocket, while the SS5 bond located towards the transmembrane domain contributes to receptor gating.
PB  - Acad Sciences Czech Republic, Inst Physiology, Prague 4
T2  - Physiological Research
T1  - Roles of Conserved Ectodomain Cysteines of the Rat P2X4 Purinoreceptor in Agonist Binding and Channel Gating
EP  - 935
IS  - 6
SP  - 927
VL  - 59
DO  - 10.33549/physiolres.931979
ER  - 

@article{
author = "Rokić, Miloš and Tvrdonova, V. and Vavra, V. and Jindrichova, M. and Obsil, T. and Stojilković, S. S. and Zemkova, H.",
year = "2010",
abstract = "Mammalian P2X receptors contain 10 conserved cysteine residues in their ectodomains, which form five disulfide bonds (SS1-5). Here, we analyzed the relevance of these disulfide pairs in rat P2X4 receptor function by replacing one or both cysteines with alanine or threonine, expressing receptors in HEK293 cells and studying their responsiveness to ATP in the absence and presence of ivermectin, an allostenic modulator of these channels. Response to ATP was not altered when both cysteines forming the SS3 bond (C132-C159) were replaced with threonines. Replacement of SS1 (C116-C165), SS2 (C126-C149) and SS4 (C217-C227), but not SS5 (C261-C270), cysteine pairs with threonines resulted in decreased sensitivity to ATP and faster deactivation times. The maximum current amplitude was reduced in SS2, SS4 and SS5 double mutants and could be partially rescued by ivermectin in SS2 and SS5 double mutants. This response pattern was also observed in numerous single residue mutants, but receptor function was not affected when the 217 cysteine was replaced with threonine or arginine or when the 261 cysteine was replaced with alanine. These results suggest that the SS1, SS2 and SS4 bonds contribute substantially to the structure of the ligand binding pocket, while the SS5 bond located towards the transmembrane domain contributes to receptor gating.",
publisher = "Acad Sciences Czech Republic, Inst Physiology, Prague 4",
journal = "Physiological Research",
title = "Roles of Conserved Ectodomain Cysteines of the Rat P2X4 Purinoreceptor in Agonist Binding and Channel Gating",
pages = "935-927",
number = "6",
volume = "59",
doi = "10.33549/physiolres.931979"
}

Rokić, M., Tvrdonova, V., Vavra, V., Jindrichova, M., Obsil, T., Stojilković, S. S.,& Zemkova, H.. (2010). Roles of Conserved Ectodomain Cysteines of the Rat P2X4 Purinoreceptor in Agonist Binding and Channel Gating. in Physiological Research
Acad Sciences Czech Republic, Inst Physiology, Prague 4., 59(6), 927-935.
https://doi.org/10.33549/physiolres.931979

Rokić M, Tvrdonova V, Vavra V, Jindrichova M, Obsil T, Stojilković SS, Zemkova H. Roles of Conserved Ectodomain Cysteines of the Rat P2X4 Purinoreceptor in Agonist Binding and Channel Gating. in Physiological Research. 2010;59(6):927-935.
doi:10.33549/physiolres.931979 .

Rokić, Miloš, Tvrdonova, V., Vavra, V., Jindrichova, M., Obsil, T., Stojilković, S. S., Zemkova, H., "Roles of Conserved Ectodomain Cysteines of the Rat P2X4 Purinoreceptor in Agonist Binding and Channel Gating" in Physiological Research, 59, no. 6 (2010):927-935,
https://doi.org/10.33549/physiolres.931979 . .