The interaction between the first transmembrane domain and the thumb of ASIC1a is critical for its N-glycosylation and trafficking.
Acid-sensing ion channel-1a (ASIC1a), the primary proton receptor in the brain, contributes to multiple diseases including stroke, epilepsy and multiple sclerosis. Thus, a better understanding of its biogenesis will provide important insights into the regulation of ASIC1a in diseases. Interestingly,...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3203923?pdf=render |
| _version_ | 1819198374993199104 |
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| author | Lan Jing Yu-Qing Jiang Qian Jiang Bin Wang Xiang-Ping Chu Xiang-Ming Zha |
| author_facet | Lan Jing Yu-Qing Jiang Qian Jiang Bin Wang Xiang-Ping Chu Xiang-Ming Zha |
| author_sort | Lan Jing |
| collection | DOAJ |
| description | Acid-sensing ion channel-1a (ASIC1a), the primary proton receptor in the brain, contributes to multiple diseases including stroke, epilepsy and multiple sclerosis. Thus, a better understanding of its biogenesis will provide important insights into the regulation of ASIC1a in diseases. Interestingly, ASIC1a contains a large, yet well organized ectodomain, which suggests the hypothesis that correct formation of domain-domain interactions at the extracellular side is a key regulatory step for ASIC1a maturation and trafficking. We tested this hypothesis here by focusing on the interaction between the first transmembrane domain (TM1) and the thumb of ASIC1a, an interaction known to be critical in channel gating. We mutated Tyr71 and Trp287, two key residues involved in the TM1-thumb interaction in mouse ASIC1a, and found that both Y71G and W287G decreased synaptic targeting and surface expression of ASIC1a. These defects were likely due to altered folding; both mutants showed increased resistance to tryptic cleavage, suggesting a change in conformation. Moreover, both mutants lacked the maturation of N-linked glycans through mid to late Golgi. These data suggest that disrupting the interaction between TM1 and thumb alters ASIC1a folding, impedes its glycosylation and reduces its trafficking. Moreover, reducing the culture temperature, an approach commonly used to facilitate protein folding, increased ASIC1a glycosylation, surface expression, current density and slowed the rate of desensitization. These results suggest that correct folding of extracellular ectodomain plays a critical role in ASIC1a biogenesis and function. |
| first_indexed | 2024-12-23T02:59:26Z |
| format | Article |
| id | doaj.art-01348445c6ff49c997ce0d34820be04c |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-23T02:59:26Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-01348445c6ff49c997ce0d34820be04c2022-12-21T18:02:27ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01610e2690910.1371/journal.pone.0026909The interaction between the first transmembrane domain and the thumb of ASIC1a is critical for its N-glycosylation and trafficking.Lan JingYu-Qing JiangQian JiangBin WangXiang-Ping ChuXiang-Ming ZhaAcid-sensing ion channel-1a (ASIC1a), the primary proton receptor in the brain, contributes to multiple diseases including stroke, epilepsy and multiple sclerosis. Thus, a better understanding of its biogenesis will provide important insights into the regulation of ASIC1a in diseases. Interestingly, ASIC1a contains a large, yet well organized ectodomain, which suggests the hypothesis that correct formation of domain-domain interactions at the extracellular side is a key regulatory step for ASIC1a maturation and trafficking. We tested this hypothesis here by focusing on the interaction between the first transmembrane domain (TM1) and the thumb of ASIC1a, an interaction known to be critical in channel gating. We mutated Tyr71 and Trp287, two key residues involved in the TM1-thumb interaction in mouse ASIC1a, and found that both Y71G and W287G decreased synaptic targeting and surface expression of ASIC1a. These defects were likely due to altered folding; both mutants showed increased resistance to tryptic cleavage, suggesting a change in conformation. Moreover, both mutants lacked the maturation of N-linked glycans through mid to late Golgi. These data suggest that disrupting the interaction between TM1 and thumb alters ASIC1a folding, impedes its glycosylation and reduces its trafficking. Moreover, reducing the culture temperature, an approach commonly used to facilitate protein folding, increased ASIC1a glycosylation, surface expression, current density and slowed the rate of desensitization. These results suggest that correct folding of extracellular ectodomain plays a critical role in ASIC1a biogenesis and function.http://europepmc.org/articles/PMC3203923?pdf=render |
| spellingShingle | Lan Jing Yu-Qing Jiang Qian Jiang Bin Wang Xiang-Ping Chu Xiang-Ming Zha The interaction between the first transmembrane domain and the thumb of ASIC1a is critical for its N-glycosylation and trafficking. PLoS ONE |
| title | The interaction between the first transmembrane domain and the thumb of ASIC1a is critical for its N-glycosylation and trafficking. |
| title_full | The interaction between the first transmembrane domain and the thumb of ASIC1a is critical for its N-glycosylation and trafficking. |
| title_fullStr | The interaction between the first transmembrane domain and the thumb of ASIC1a is critical for its N-glycosylation and trafficking. |
| title_full_unstemmed | The interaction between the first transmembrane domain and the thumb of ASIC1a is critical for its N-glycosylation and trafficking. |
| title_short | The interaction between the first transmembrane domain and the thumb of ASIC1a is critical for its N-glycosylation and trafficking. |
| title_sort | interaction between the first transmembrane domain and the thumb of asic1a is critical for its n glycosylation and trafficking |
| url | http://europepmc.org/articles/PMC3203923?pdf=render |
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