Immobilization of arrestin-3 on different biosensor platforms for evaluating GPCR binding

Avsar, Saziye Yorulmaz and Kapinos, Larisa E. and Schoenenberger, Cora-Ann and Schertler, Gebhard F. X. and Muhle, Jonas and Meger, Benoit and Lim, Roderick Y. H. and Ostermaier, Martin K. and Lesca, Elena and Palivan, Cornelia G.. (2020) Immobilization of arrestin-3 on different biosensor platforms for evaluating GPCR binding. Physical Chemistry Chemical Physics, 22 (41). pp. 24086-24096.

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G protein-coupled receptors (GPCRs) are a large and ubiquitous family of membrane receptors of great pharmacological interest. Cell-based assays are the primary tool for assessing GPCR interactions and activation but their design and intrinsic complexity limit their application. Biosensor-based assays that directly and specifically report GPCR-protein binding (e.g. arrestin or G protein) could provide a good alternative. We present an approach based on the stable immobilization of different arrestin-3 proteins (wild type, and two mutants, mutant X (arrestin-3 I386A) and mutant Y (arrestin-3 R393E)) via histidine tags on NTA(Ni2+)-coated sensors in a defined orientation. Using biolayer interferometry (BLI), surface plasmon resonance (SPR), and quartz crystal microbalance with dissipation (QCM-D), we were able to follow the interaction between the different arrestin-3 proteins and a representative GPCR, jumping spider rhodopsin-1 (JSR1), in a label-free manner in real-time. The interactions were quantified as binding affinity, association and dissociation rate constants. The combination of surface-based biosensing methods indicated that JSR1 showed the strongest binding to arrestin mutant Y. Taken together, this work introduces direct label-free, biosensor-based screening approaches that can be easily adapted for testing interactions of proteins and other compounds with different GPCRs.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Nanobiology Argovia (Lim)
05 Faculty of Science > Departement Chemie > Chemie > Physikalische Chemie (Palivan)
UniBasel Contributors:Yorulmaz Avsar, Saziye and Kapinos Schneider, Larisa E. E and Schönenberger, Cora-Ann and Lim, Roderick Y.H. and Palivan, Cornelia G
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Royal Society of Chemistry
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:11 Mar 2021 11:44
Deposited On:11 Mar 2021 11:44

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