A genetically encoded nanobody sensor reveals conformational diversity in β-arrestins orchestrated by distinct seven transmembrane receptors
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F25%3A00639402" target="_blank" >RIV/61388963:_____/25:00639402 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11110/25:10503588
Result on the web
<a href="https://doi.org/10.1073/pnas.2507384122" target="_blank" >https://doi.org/10.1073/pnas.2507384122</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1073/pnas.2507384122" target="_blank" >10.1073/pnas.2507384122</a>
Alternative languages
Result language
angličtina
Original language name
A genetically encoded nanobody sensor reveals conformational diversity in β-arrestins orchestrated by distinct seven transmembrane receptors
Original language description
Agonist-induced interaction of G protein-coupled receptors (GPCRs) with beta- arrestins (beta arrs) is a critical mechanism that regulates the spatiotemporal pattern of receptor localization and signaling. While the underlying mechanism governing GPCR-beta arr interaction is primarily conserved and involves receptor activation and phosphorylation, there are several examples of receptor-specific fine-tuning of beta arr-mediated functional outcomes. Considering the key contribution of conformational plasticity of beta arrs in driving receptor-specific functional responses, it is important to develop novel sensors capable of reporting distinct beta arr conformations in cellular context. Here, we design an intrabody version of a beta arr-recognizing nanobody (nanobody32), referred to as intrabody32 (Ib32), in NanoLuc enzyme complementation assay format and measure its ability to recognize beta arr1 and 2 in live cells upon activation of a broad set of GPCRs. Ib32 robustly recognizes activated beta arr1 and 2 in the plasma membrane and endosomes, and effectively mirrors beta arr recruitment profile upon stimulation of selected GPCRs. We also design an Ib32 sensor for polarization microscopy with a change in linear dichroism as readout and demonstrate its utility for monitoring beta arr activation upon stimulation of selected GPCRs by natural and biased agonists. Taken together with a previously described sensor of beta arr1 activation, Ib32 underscores the inherent flexibility encoded in beta arrs and conformational diversity imparted by different GPCRs, which is further corroborated using an orthogonal limited proteolysis assay. Our study presents Ib32 as a sensor of beta arr activation and highlights the structural diversity of beta arrs, which likely allows their ability to interact with, and regulate, a large repertoire of GPCRs.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10610 - Biophysics
Result continuities
Project
<a href="/en/project/LTAIN19167" target="_blank" >LTAIN19167: Visualizing GPCR-G-protein-beta-arrestin supercomplexes in living cells using two-photon polarization microscopy</a><br>
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2025
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Proceedings of the National Academy of Sciences of the United States of America
ISSN
0027-8424
e-ISSN
1091-6490
Volume of the periodical
122
Issue of the periodical within the volume
37
Country of publishing house
US - UNITED STATES
Number of pages
12
Pages from-to
e2507384122
UT code for WoS article
001576021100001
EID of the result in the Scopus database
2-s2.0-105015671723