A novel nanobody-based bio-assay using functional complementation of a split nanoluciferase to monitor Mu- opioid receptor activation
- Author
- Lakshmi Vasudevan and Christophe Stove (UGent)
- Organization
- Project
- Abstract
- The Mu opioid receptor (MOR) has been the subject of intense research over the past decades, especially in the field of analgesic therapeutics. It is the primary target for both clinical and recreational opioids. Recently, camelid-derived nanobodies have received significant attention due to their applicability in stabilizing the crystal structure of activated MOR, via specific recognition of and binding to the active receptor conformation. In the present study, we developed and applied a novel bio-assay to monitor MOR activation, utilizing intracellular expression of one such nanobody, Nb39. The principle of functional complementation of a split nanoluciferase was used to assess recruitment of Nb39 to MOR, following activation by a set of five synthetic opioids. The obtained pharmacological parameters—negative logarithm of EC50 (pEC50, as a measure of potency) and maximal response provoked by a ligand (Emax, as a measure of efficacy; relative to hydromorphone)—were compared with those obtained using a G protein recruitment assay, in which a mini-Gi protein (engineered GTPase domain of Gαi subunit) is recruited to activated MOR. Similar EC50 but distinct Emax values were obtained with both bio-assays, with lower Emax values for the Nbbased bio-assay. Both bio-assays may assist to gain better insight into activation of the MOR.
- Keywords
- New psychoactive substances, Mu opioid receptor, Nanobody, Bio-assay, Functional complementation, Nanoluciferase
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-8687069
- MLA
- Vasudevan, Lakshmi, and Christophe Stove. “A Novel Nanobody-Based Bio-Assay Using Functional Complementation of a Split Nanoluciferase to Monitor Mu- Opioid Receptor Activation.” ANALYTICAL AND BIOANALYTICAL CHEMISTRY, vol. 412, no. 29, 2020, pp. 8015–22, doi:10.1007/s00216-020-02945-6.
- APA
- Vasudevan, L., & Stove, C. (2020). A novel nanobody-based bio-assay using functional complementation of a split nanoluciferase to monitor Mu- opioid receptor activation. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 412(29), 8015–8022. https://doi.org/10.1007/s00216-020-02945-6
- Chicago author-date
- Vasudevan, Lakshmi, and Christophe Stove. 2020. “A Novel Nanobody-Based Bio-Assay Using Functional Complementation of a Split Nanoluciferase to Monitor Mu- Opioid Receptor Activation.” ANALYTICAL AND BIOANALYTICAL CHEMISTRY 412 (29): 8015–22. https://doi.org/10.1007/s00216-020-02945-6.
- Chicago author-date (all authors)
- Vasudevan, Lakshmi, and Christophe Stove. 2020. “A Novel Nanobody-Based Bio-Assay Using Functional Complementation of a Split Nanoluciferase to Monitor Mu- Opioid Receptor Activation.” ANALYTICAL AND BIOANALYTICAL CHEMISTRY 412 (29): 8015–8022. doi:10.1007/s00216-020-02945-6.
- Vancouver
- 1.Vasudevan L, Stove C. A novel nanobody-based bio-assay using functional complementation of a split nanoluciferase to monitor Mu- opioid receptor activation. ANALYTICAL AND BIOANALYTICAL CHEMISTRY. 2020;412(29):8015–22.
- IEEE
- [1]L. Vasudevan and C. Stove, “A novel nanobody-based bio-assay using functional complementation of a split nanoluciferase to monitor Mu- opioid receptor activation,” ANALYTICAL AND BIOANALYTICAL CHEMISTRY, vol. 412, no. 29, pp. 8015–8022, 2020.
@article{8687069, abstract = {{The Mu opioid receptor (MOR) has been the subject of intense research over the past decades, especially in the field of analgesic therapeutics. It is the primary target for both clinical and recreational opioids. Recently, camelid-derived nanobodies have received significant attention due to their applicability in stabilizing the crystal structure of activated MOR, via specific recognition of and binding to the active receptor conformation. In the present study, we developed and applied a novel bio-assay to monitor MOR activation, utilizing intracellular expression of one such nanobody, Nb39. The principle of functional complementation of a split nanoluciferase was used to assess recruitment of Nb39 to MOR, following activation by a set of five synthetic opioids. The obtained pharmacological parameters—negative logarithm of EC50 (pEC50, as a measure of potency) and maximal response provoked by a ligand (Emax, as a measure of efficacy; relative to hydromorphone)—were compared with those obtained using a G protein recruitment assay, in which a mini-Gi protein (engineered GTPase domain of Gαi subunit) is recruited to activated MOR. Similar EC50 but distinct Emax values were obtained with both bio-assays, with lower Emax values for the Nbbased bio-assay. Both bio-assays may assist to gain better insight into activation of the MOR.}}, author = {{Vasudevan, Lakshmi and Stove, Christophe}}, issn = {{1618-2642}}, journal = {{ANALYTICAL AND BIOANALYTICAL CHEMISTRY}}, keywords = {{New psychoactive substances,Mu opioid receptor,Nanobody,Bio-assay,Functional complementation,Nanoluciferase}}, language = {{eng}}, number = {{29}}, pages = {{8015--8022}}, title = {{A novel nanobody-based bio-assay using functional complementation of a split nanoluciferase to monitor Mu- opioid receptor activation}}, url = {{http://doi.org/10.1007/s00216-020-02945-6}}, volume = {{412}}, year = {{2020}}, }
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