Altered allostery of the left flipper domain underlies the weak ATP response of rat P2X5 receptors
| Autor: | Yan Liu, Li-Dong Huang, Heng-Shan Wang, Liang-Fei Sun, Ye Yu, Hong Liang, Ying-Zhe Fan, Michael X. Zhu, Chang-Run Guo, Yang Yang, Jin Wang, Xiaoyang Cheng, Yun Tian |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Recombinant Fusion Proteins Transgene Allosteric regulation Gating Molecular Dynamics Simulation Biochemistry 03 medical and health sciences Adenosine Triphosphate Protein Domains Cations medicine Extracellular Animals Humans Biotinylation Disulfides Receptor Molecular Biology 030102 biochemistry & molecular biology Chemistry Purinergic receptor Skeletal muscle Exons Cell Biology Rats Cell biology Transmembrane domain Cross-Linking Reagents HEK293 Cells 030104 developmental biology medicine.anatomical_structure Receptors Purinergic P2X5 Allosteric Site Molecular Biophysics |
| Zdroj: | J Biol Chem |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.ra119.009959 |
| Popis: | Although the extracellular ATP-gated cation channel purinergic receptor P2X5 is widely expressed in heart, skeletal muscle, and immune and nervous systems in mammals, little is known about its functions and channel-gating activities. This lack of knowledge is due to P2X5's weak ATP responses in several mammalian species, such as humans, rats, and mice. WT human P2X5 (hP2X5(Δ328–349)) does not respond to ATP, whereas a full-length variant, hP2X5 (hP2X5-FL), containing exon 10 encoding the second hP2X5 transmembrane domain (TM2), does. However, although rat P2X5 (rP2X5) has a full-length TM2, ATP induces only weak currents in rP2X5, which prompted us to investigate the mechanism underlying this small ATP response. Here, we show that single replacements of specific rP2X5 residues with the corresponding residues in hP2X5 (S191F or F195H) significantly enhance the current amplitude of rP2X5. Using a combination of engineered disulfide cross-linking, single-channel recording, and molecular modeling, we interrogated the effects of S191F and F195H substitutions on the allostery of the left flipper (LF) domain. On the basis of our findings, we propose that the bound ATP-induced distinct allostery of the LF domain with that of other functional subtypes has caused the weak ATP response of rP2X5 receptors. The findings of our study provide the prerequisite for future transgenic studies on the physiological and pathological functions of P2X5 receptors. |
| Databáze: | OpenAIRE |
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