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Rapid prediction of G protein-coupled receptor structures using nanoluciferase assay
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Yu-ming ZHUANG1, 2, Lu-lu GUO2, Guo-xing FANG1, 2, Xin LUO2, Si-yuan SHEN2, Fan YANG2, *, Jiu-yao ZHOU1, *
Acta Pharmaceutica Sinica | 2023, 58(5) : 1267 - 1274
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Acta Pharmaceutica Sinica | 2023, 58(5): 1267-1274
• Original Articles •
Rapid prediction of G protein-coupled receptor structures using nanoluciferase assay
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Yu-ming ZHUANG1, 2, Lu-lu GUO2, Guo-xing FANG1, 2, Xin LUO2, Si-yuan SHEN2, Fan YANG2, *, Jiu-yao ZHOU1, *
Affiliations
  • 1. School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 511400, China
  • 2. Shandong University School of Basic Medicine, Jinan 250014, China
Published: 2023-05-12 doi: 10.16438/j.0513-4870.2023-0076
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Using beta-2 adrenergic receptor, 5-hydroxytryptamine and angiotensin Ⅱtype 1 receptor as control, we here established a method for rapid prediction of the initial position amino acids of N-terminal, C-terminal, intracellular loops, extracellular loops and transmembrane (TM) regions in G protein-coupled receptors (GPCRs), and successfully predicted the structure of Mas-related G protein-coupled receptors X3 (MRGPRX3). To achieve this purpose, nanoluciferase (Nluc) was inserted into the different sites of these GPCRs′ sequence by sequence and ligation-independent cloning (SLIC) method, and the luminescence value were measured to distinguish the different parts of GPCRs. The results showed that luminescence values of NLuc luciferase at TM region were less than 100 000, and the values were higher than 1 000 000 at N terminal, C terminal, or extracellular loops and intracellular loops, and the values were between 100 000 and 500 000 at junction. The predicted MRGPRX3 structure was analyzed in detail and was compared with AlphaFold predicted structure. In conclusion, this method could provide useful information of GPCR structure model for the ligand virtual screening, and could provide certain experimental basis for structural pharmacology.

G protein-coupled receptor  /  nanoluciferase  /  flexible area  /  rigid area  /  prediction of structure
Yu-ming ZHUANG, Lu-lu GUO, Guo-xing FANG, Xin LUO, Si-yuan SHEN, Fan YANG, Jiu-yao ZHOU. Rapid prediction of G protein-coupled receptor structures using nanoluciferase assay[J]. Acta Pharmaceutica Sinica, 2023 , 58 (5) : 1267 -1274 . DOI: 10.16438/j.0513-4870.2023-0076
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Article Info
doi: 10.16438/j.0513-4870.2023-0076
  • Receive Date：2023-01-29
  • Online Date：2025-11-21
  • Published：2023-05-12
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History
  • Received：2023-01-29
  • Revised：2023-02-20
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Affiliations
    1. School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 511400, China
    2. Shandong University School of Basic Medicine, Jinan 250014, China
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