Real-time visualization of ligand-specific conformational dynamics of GPCR C-terminal domain in living cells Article Swipe
Lin Zhao
,
Xinyao Chen
,
Lijuan Ma
,
Hang Fu
,
Hao Wang
,
Kai Yang
,
Zijian Li
,
Fangfu Ye
,
Ying Lü
,
Shuxin Hu
,
Xiaogang Li
·
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1101/2025.07.17.665247
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1101/2025.07.17.665247
The intrinsically disordered C-terminal domain (CTD) serves as a critical regulatory element in GPCR 1–3 . However, directly interrogating the CTD responses to different ligands is challenging due to its high flexibility, which renders it invisible to conventional structural biology techniques. To address the challenge, we developed a live-cell fluorescence imaging strategy that enables real-time visualization of CTD conformational transitions under physiological conditions. Our dual-mode approach integrates single-cell fluorescence lifetime imaging microscopy (FLIM) with single-molecule total internal reflection fluorescence microscopy (TIRFM), facilitating multi-scale analysis. Our data revealed that the dynamics of both full-length and truncated CTDs in the β2-adrenergic receptors are ligand-specific. By bridging single-molecule dynamics with ensemble cellular responses, our method uncovered previously inaccessible molecular mechanisms underlying receptor activation. This advance not only elucidates how GPCRs transduce ligand binding into functional outcomes but also establishes a versatile platform for drug discovery, enabling rapid assessment of ligand efficacy and receptor activity in physiological contexts.
Related Topics
Concepts
G protein-coupled receptor
Terminal (telecommunication)
Domain (mathematical analysis)
Visualization
Dynamics (music)
Ligand (biochemistry)
Chemistry
Biophysics
Computer science
Cell biology
Computational biology
Biology
Receptor
Physics
Data mining
Biochemistry
Mathematics
Computer network
Acoustics
Mathematical analysis
Metadata
- Type
- preprint
- Language
- en
- Landing Page
- https://doi.org/10.1101/2025.07.17.665247
- https://www.biorxiv.org/content/biorxiv/early/2025/07/21/2025.07.17.665247.full.pdf
- OA Status
- green
- References
- 73
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4412528621
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4412528621Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1101/2025.07.17.665247Digital Object Identifier
- Title
-
Real-time visualization of ligand-specific conformational dynamics of GPCR C-terminal domain in living cellsWork title
- Type
-
preprintOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2025Year of publication
- Publication date
-
2025-07-21Full publication date if available
- Authors
-
Lin Zhao, Xinyao Chen, Lijuan Ma, Hang Fu, Hao Wang, Kai Yang, Zijian Li, Fangfu Ye, Ying Lü, Shuxin Hu, Xiaogang LiList of authors in order
- Landing page
-
https://doi.org/10.1101/2025.07.17.665247Publisher landing page
- PDF URL
-
https://www.biorxiv.org/content/biorxiv/early/2025/07/21/2025.07.17.665247.full.pdfDirect link to full text PDF
- Open access
-
YesWhether a free full text is available
- OA status
-
greenOpen access status per OpenAlex
- OA URL
-
https://www.biorxiv.org/content/biorxiv/early/2025/07/21/2025.07.17.665247.full.pdfDirect OA link when available
- Concepts
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G protein-coupled receptor, Terminal (telecommunication), Domain (mathematical analysis), Visualization, Dynamics (music), Ligand (biochemistry), Chemistry, Biophysics, Computer science, Cell biology, Computational biology, Biology, Receptor, Physics, Data mining, Biochemistry, Mathematics, Computer network, Acoustics, Mathematical analysisTop concepts (fields/topics) attached by OpenAlex
- Cited by
-
0Total citation count in OpenAlex
- References (count)
-
73Number of works referenced by this work
- Related works (count)
-
10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.advance | 122 |
| abstract_inverted_index.binding | 130 |
| abstract_inverted_index.biology | 40 |
| abstract_inverted_index.element | 12 |
| abstract_inverted_index.enables | 54 |
| abstract_inverted_index.imaging | 51, 71 |
| abstract_inverted_index.ligands | 25 |
| abstract_inverted_index.renders | 34 |
| abstract_inverted_index.(TIRFM), | 81 |
| abstract_inverted_index.Abstract | 0 |
| abstract_inverted_index.However, | 17 |
| abstract_inverted_index.activity | 151 |
| abstract_inverted_index.approach | 66 |
| abstract_inverted_index.bridging | 104 |
| abstract_inverted_index.cellular | 109 |
| abstract_inverted_index.critical | 10 |
| abstract_inverted_index.directly | 18 |
| abstract_inverted_index.dynamics | 90, 106 |
| abstract_inverted_index.efficacy | 148 |
| abstract_inverted_index.enabling | 143 |
| abstract_inverted_index.ensemble | 108 |
| abstract_inverted_index.internal | 77 |
| abstract_inverted_index.lifetime | 70 |
| abstract_inverted_index.outcomes | 133 |
| abstract_inverted_index.platform | 139 |
| abstract_inverted_index.receptor | 119, 150 |
| abstract_inverted_index.revealed | 87 |
| abstract_inverted_index.strategy | 52 |
| abstract_inverted_index.analysis. | 84 |
| abstract_inverted_index.contexts. | 154 |
| abstract_inverted_index.developed | 47 |
| abstract_inverted_index.different | 24 |
| abstract_inverted_index.dual-mode | 65 |
| abstract_inverted_index.invisible | 36 |
| abstract_inverted_index.live-cell | 49 |
| abstract_inverted_index.molecular | 116 |
| abstract_inverted_index.real-time | 55 |
| abstract_inverted_index.receptors | 100 |
| abstract_inverted_index.responses | 22 |
| abstract_inverted_index.transduce | 128 |
| abstract_inverted_index.truncated | 95 |
| abstract_inverted_index.uncovered | 113 |
| abstract_inverted_index.versatile | 138 |
| abstract_inverted_index.C-terminal | 4 |
| abstract_inverted_index.assessment | 145 |
| abstract_inverted_index.challenge, | 45 |
| abstract_inverted_index.discovery, | 142 |
| abstract_inverted_index.disordered | 3 |
| abstract_inverted_index.elucidates | 125 |
| abstract_inverted_index.functional | 132 |
| abstract_inverted_index.integrates | 67 |
| abstract_inverted_index.mechanisms | 117 |
| abstract_inverted_index.microscopy | 72, 80 |
| abstract_inverted_index.previously | 114 |
| abstract_inverted_index.reflection | 78 |
| abstract_inverted_index.regulatory | 11 |
| abstract_inverted_index.responses, | 110 |
| abstract_inverted_index.structural | 39 |
| abstract_inverted_index.underlying | 118 |
| abstract_inverted_index.activation. | 120 |
| abstract_inverted_index.challenging | 27 |
| abstract_inverted_index.conditions. | 63 |
| abstract_inverted_index.establishes | 136 |
| abstract_inverted_index.full-length | 93 |
| abstract_inverted_index.multi-scale | 83 |
| abstract_inverted_index.single-cell | 68 |
| abstract_inverted_index.techniques. | 41 |
| abstract_inverted_index.transitions | 60 |
| abstract_inverted_index.conventional | 38 |
| abstract_inverted_index.facilitating | 82 |
| abstract_inverted_index.flexibility, | 32 |
| abstract_inverted_index.fluorescence | 50, 69, 79 |
| abstract_inverted_index.inaccessible | 115 |
| abstract_inverted_index.interrogating | 19 |
| abstract_inverted_index.intrinsically | 2 |
| abstract_inverted_index.physiological | 62, 153 |
| abstract_inverted_index.visualization | 56 |
| abstract_inverted_index.conformational | 59 |
| abstract_inverted_index.β2-adrenergic | 99 |
| abstract_inverted_index.single-molecule | 75, 105 |
| abstract_inverted_index.ligand-specific. | 102 |
| cited_by_percentile_year | |
| corresponding_author_ids | https://openalex.org/A5100355322, https://openalex.org/A5064133005, https://openalex.org/A5015934449 |
| countries_distinct_count | 1 |
| institutions_distinct_count | 11 |
| corresponding_institution_ids | https://openalex.org/I19820366, https://openalex.org/I4210114608, https://openalex.org/I4210159876, https://openalex.org/I4210165038 |
| citation_normalized_percentile.value | 0.29180175 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | False |