A new MAS-related G protein-coupled receptor X2 cell membrane chromatography analysis model based on HALO-tag technology and its applications
Copyright © 2023 Elsevier B.V. All rights reserved..
Cell membrane chromatography (CMC) is an effective method for studying receptors with multiple transmembrane structure such as MAS-related G protein-coupled receptor X2 (MrgX2). CMC relies on the maintenance of the complete biological structure of a membrane receptor; however, it needs to be further improved to obtain a more convenient and stable CMC model. In the present study, the haloalkane dehalogenase protein tag (HALO-tag) technology was used to construct a new MrgX2/CMC model. The fusion receptors of MrgX2 with HALO-tag at the C terminus were expressed in HEK293 cells. The silica gel was modified with a substrate of HALO-tag (chloroalkanes) via one-step acylation for the rapid capture of fusion receptors. The new CMC model (MrgX2-HALO-tag/CMC model) was not only quicker to prepare but also more stable and had a longer lifespan than a previous MrgX2-SNAP-tag/CMC model. In combination with the high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) system, the MrgX2-HALO-tag/CMC model was used to screen and identify bioactive components in traditional Chinese medicine. Using this combination, sanggenon C and morusin were identified from Mori Cortex as anti-pseudo-allergic components. The MrgX2-HALO-tag/CMC model alone was also applied to analyze ligand-receptor interaction. The affinity order of four ligands to MrgX2 was as follows: desipramine < imipramine < amitriptyline < clomipramine. This was consistent with the results obtained using the MrgX2-SNAP-tag/CMC model. The MrgX2-HALO-tag/CMC model provides ideas and application prospects for the immobilization of cell membrane that contains receptors with more transmembrane structures.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 2023 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:268 |
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| Enthalten in: |
Talanta - 268(2023), Pt 1 vom: 01. Feb., Seite 125317 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Jia, Qianqian [VerfasserIn] |
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| Links: |
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| Themen: |
Cell membrane chromatography |
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| Anmerkungen: |
Date Completed 27.11.2023 Date Revised 27.11.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.talanta.2023.125317 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM363714146 |
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| 100 | 1 | |a Jia, Qianqian |e verfasserin |4 aut | |
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| 520 | |a Copyright © 2023 Elsevier B.V. All rights reserved. | ||
| 520 | |a Cell membrane chromatography (CMC) is an effective method for studying receptors with multiple transmembrane structure such as MAS-related G protein-coupled receptor X2 (MrgX2). CMC relies on the maintenance of the complete biological structure of a membrane receptor; however, it needs to be further improved to obtain a more convenient and stable CMC model. In the present study, the haloalkane dehalogenase protein tag (HALO-tag) technology was used to construct a new MrgX2/CMC model. The fusion receptors of MrgX2 with HALO-tag at the C terminus were expressed in HEK293 cells. The silica gel was modified with a substrate of HALO-tag (chloroalkanes) via one-step acylation for the rapid capture of fusion receptors. The new CMC model (MrgX2-HALO-tag/CMC model) was not only quicker to prepare but also more stable and had a longer lifespan than a previous MrgX2-SNAP-tag/CMC model. In combination with the high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) system, the MrgX2-HALO-tag/CMC model was used to screen and identify bioactive components in traditional Chinese medicine. Using this combination, sanggenon C and morusin were identified from Mori Cortex as anti-pseudo-allergic components. The MrgX2-HALO-tag/CMC model alone was also applied to analyze ligand-receptor interaction. The affinity order of four ligands to MrgX2 was as follows: desipramine < imipramine < amitriptyline < clomipramine. This was consistent with the results obtained using the MrgX2-SNAP-tag/CMC model. The MrgX2-HALO-tag/CMC model provides ideas and application prospects for the immobilization of cell membrane that contains receptors with more transmembrane structures | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Cell membrane chromatography | |
| 650 | 4 | |a HALO-tag technology | |
| 650 | 4 | |a HPLC-MS/MS | |
| 650 | 4 | |a MrgX2 receptor | |
| 650 | 4 | |a Pseudo-allergic compounds | |
| 650 | 7 | |a Nerve Tissue Proteins |2 NLM | |
| 650 | 7 | |a Receptors, Neuropeptide |2 NLM | |
| 650 | 7 | |a Receptors, G-Protein-Coupled |2 NLM | |
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| 700 | 1 | |a Miao, Chenyang |e verfasserin |4 aut | |
| 700 | 1 | |a Feng, Jingting |e verfasserin |4 aut | |
| 700 | 1 | |a Ding, Yifan |e verfasserin |4 aut | |
| 700 | 1 | |a Zhou, Tongpei |e verfasserin |4 aut | |
| 700 | 1 | |a Han, Shengli |e verfasserin |4 aut | |
| 700 | 1 | |a He, Langchong |e verfasserin |4 aut | |
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