A conserved residue in the P2X4 receptor has a nonconserved function in ATP recognition
Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved..
Highly conserved amino acids are generally anticipated to have similar functions across a protein superfamily, including that of the P2X ion channels, which are gated by extracellular ATP. However, whether and how these functions are conserved becomes less clear when neighboring amino acids are not conserved. Here, we investigate one such case, focused on the highly conserved residue from P2X4, E118 (rat P2X4 numbering, rP2X4), a P2X subtype associated with human neuropathic pain. When we compared the crystal structures of P2X4 with those of other P2X subtypes, including P2X3, P2X7, and AmP2X, we observed a slightly altered side-chain orientation of E118. We used protein chimeras, double-mutant cycle analysis, and molecular modeling to reveal that E118 forms specific contacts with amino acids in the "beak" region, which facilitates ATP binding to rP2X4. These contacts are not present in other subtypes because of sequence variance in the beak region, resulting in decoupling of this conserved residue from ATP recognition and/or channel gating of P2X receptors. Our study provides an example of a conserved residue with a specific role in functional proteins enabled by adjacent nonconserved residues. The unique role established by the E118-beak region contact provides a blueprint for the development of subtype-specific inhibitors of P2X4.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2021 |
|---|---|
| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:296 |
|---|---|
| Enthalten in: |
The Journal of biological chemistry - 296(2021) vom: 05. Jan., Seite 100655 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Chen, Ping-Fang [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 23.08.2021 Date Revised 02.04.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1016/j.jbc.2021.100655 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM324549822 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM324549822 | ||
| 003 | DE-627 | ||
| 005 | 20240402232709.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2021 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1016/j.jbc.2021.100655 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1360.xml |
| 035 | |a (DE-627)NLM324549822 | ||
| 035 | |a (NLM)33901491 | ||
| 035 | |a (PII)S0021-9258(21)00442-7 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Chen, Ping-Fang |e verfasserin |4 aut | |
| 245 | 1 | 2 | |a A conserved residue in the P2X4 receptor has a nonconserved function in ATP recognition |
| 264 | 1 | |c 2021 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 23.08.2021 | ||
| 500 | |a Date Revised 02.04.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved. | ||
| 520 | |a Highly conserved amino acids are generally anticipated to have similar functions across a protein superfamily, including that of the P2X ion channels, which are gated by extracellular ATP. However, whether and how these functions are conserved becomes less clear when neighboring amino acids are not conserved. Here, we investigate one such case, focused on the highly conserved residue from P2X4, E118 (rat P2X4 numbering, rP2X4), a P2X subtype associated with human neuropathic pain. When we compared the crystal structures of P2X4 with those of other P2X subtypes, including P2X3, P2X7, and AmP2X, we observed a slightly altered side-chain orientation of E118. We used protein chimeras, double-mutant cycle analysis, and molecular modeling to reveal that E118 forms specific contacts with amino acids in the "beak" region, which facilitates ATP binding to rP2X4. These contacts are not present in other subtypes because of sequence variance in the beak region, resulting in decoupling of this conserved residue from ATP recognition and/or channel gating of P2X receptors. Our study provides an example of a conserved residue with a specific role in functional proteins enabled by adjacent nonconserved residues. The unique role established by the E118-beak region contact provides a blueprint for the development of subtype-specific inhibitors of P2X4 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a channel gating | |
| 650 | 4 | |a electrophysiology | |
| 650 | 4 | |a ion channel | |
| 650 | 4 | |a molecular dynamics | |
| 650 | 4 | |a purinergic receptor | |
| 650 | 4 | |a side-chain orientation | |
| 650 | 4 | |a structure–function | |
| 650 | 7 | |a Receptors, Purinergic P2X4 |2 NLM | |
| 650 | 7 | |a Adenosine Triphosphate |2 NLM | |
| 650 | 7 | |a 8L70Q75FXE |2 NLM | |
| 700 | 1 | |a Ma, Xue-Fei |e verfasserin |4 aut | |
| 700 | 1 | |a Sun, Liang-Fei |e verfasserin |4 aut | |
| 700 | 1 | |a Tian, Yun |e verfasserin |4 aut | |
| 700 | 1 | |a Fan, Ying-Zhe |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Peiwang |e verfasserin |4 aut | |
| 700 | 1 | |a Xiao, Zhihong |e verfasserin |4 aut | |
| 700 | 1 | |a Zhu, Michael X |e verfasserin |4 aut | |
| 700 | 1 | |a Guo, Chang-Run |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Changzhu |e verfasserin |4 aut | |
| 700 | 1 | |a Yu, Ye |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Jin |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t The Journal of biological chemistry |d 1945 |g 296(2021) vom: 05. Jan., Seite 100655 |w (DE-627)NLM000004995 |x 1083-351X |7 nnns |
| 773 | 1 | 8 | |g volume:296 |g year:2021 |g day:05 |g month:01 |g pages:100655 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1016/j.jbc.2021.100655 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 296 |j 2021 |b 05 |c 01 |h 100655 | ||