气候变化领域集成服务门户(CCPortal): Localization of the gate and selectivity filter of the full-length P2X7 receptor

CCPortal

DOI	10.1073/pnas.1610414114
	Localization of the gate and selectivity filter of the full-length P2X7 receptor
	Pippel A.; Stolz M.; Woltersdorf R.; Kless A.; Schmalzing G.; Markwardt F.
发表日期	2017
ISSN	0027-8424
起始页码	E2156
结束页码	E2165
卷号	114 期号:11
英文摘要	The P2X7 receptor (P2X7R) belongs to the P2X family of ATP-gated cation channels. P2X7Rs are expressed in epithelial cells, leukocytes, and microglia, and they play important roles in immunological and inflammatory processes. P2X7Rs are obligate homotrimers, with each subunit having two transmembrane helices, TM1 and TM2. Structural and functional data regarding the P2X2 and P2X4 receptors indicate that the central trihelical TM2 bundle forms the intrinsic transmembrane channel of P2X receptors. Here, we studied the accessibility of single cysteines substituted along the pre-TM2 and TM2 helix (residues 327-357) of the P2X7R using as readouts (i) the covalent maleimide fluorescence accessibility of the surface-bound P2X7R and (ii) covalent modulation of macroscopic and single-channel currents using extracellularly and intracellularly applied methanethiosulfonate (MTS) reagents.We found that the channel opening extends from the pre-TM2 region through the outer half of the trihelical TM2 channel. Covalently adducted MTS ethylammonium+ (MTSEA+) strongly increased the probability that the channel was open by delaying channel closing of seven of eight responsive human P2X7R (hP2X7R) mutants. Structural modeling, as supported by experimental probing, suggested that resulting intraluminal hydrogen bonding interactions stabilize the open-channel state. The additional decrease in singlechannel conductance by MTSEA+ in five of seven positions identified Y336, S339, L341C, Y343, and G345 as the narrowest part of the channel lumen. The gate and ion-selectivity filter of the P2X7R could be colocalized at and around residue S342. None of our results provided any evidence for dilation of the hP2X7R channel on sustained stimulation with ATP4.
英文关键词	Cysteine-Scanning Accessibility Mutagenesis; P2X7 receptor; P2X7 Receptor Homology Model; Single-Channel Conductance; Single-Channel Open Probability
语种	英语
scopus关键词	cysteine; hydrogen; maleimide; purinergic P2X7 receptor; adenosine triphosphate; carbocyanine; cyanine dye 5; purinergic P2X7 receptor; Article; fluorescence; hydrogen bond; modulation; priority journal; amino acid substitution; channel gating; chemistry; genetics; metabolism; molecular model; protein conformation; protein transport; structure activity relation; Adenosine Triphosphate; Amino Acid Substitution; Carbocyanines; Cysteine; Hydrogen Bonding; Ion Channel Gating; Models, Molecular; Protein Conformation; Protein Transport; Receptors, Purinergic P2X7; Structure-Activity Relationship
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/160655
作者单位	Pippel, A., Julius Bernstein Institute for Physiology, Martin Luther University, Halle/Saale, D-06097, Germany; Stolz, M., Molecular Pharmacology, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, D-52074, Germany; Woltersdorf, R., Molecular Pharmacology, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, D-52074, Germany; Kless, A., Grönenthal Innovation, Department of Drug Discovery Technologies, Grönenthal GmbH, Aachen, D-52078, Germany; Schmalzing, G., Molecular Pharmacology, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, D-52074, Germany; Markwardt, F., Julius Bernstein Institute for Physiology, Martin Luther University, Halle/Saale, D-06097, Germany
推荐引用方式 GB/T 7714	Pippel A.,Stolz M.,Woltersdorf R.,et al. Localization of the gate and selectivity filter of the full-length P2X7 receptor[J],2017,114(11).
APA	Pippel A.,Stolz M.,Woltersdorf R.,Kless A.,Schmalzing G.,&Markwardt F..(2017).Localization of the gate and selectivity filter of the full-length P2X7 receptor.Proceedings of the National Academy of Sciences of the United States of America,114(11).
MLA	Pippel A.,et al."Localization of the gate and selectivity filter of the full-length P2X7 receptor".Proceedings of the National Academy of Sciences of the United States of America 114.11(2017).