1. Protocol for identifying physiologically relevant binding proteins of G-protein-coupled receptors.
- Author
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Huang BX and Kim HY
- Subjects
- Cell Membrane, Immunoprecipitation, Mass Spectrometry, Carrier Proteins, Receptors, G-Protein-Coupled genetics
- Abstract
G-protein-coupled receptors (GPCRs) are important therapeutic targets expressed on the cell surface. Here, we present a protocol for identifying physiologically relevant binding proteins of adhesion GPCR GPR110. We describe steps for in-cell chemical crosslinking, immunoprecipitation, and quantitative high-resolution mass spectrometry. Notably, we detail a label-free quantitation strategy that eliminates irrelevant interacting proteins using an inactive GPR110 mutant with impaired surface expression. Furthermore, we outline procedures for validating the identified partners. For complete details on the use and execution of this protocol, please refer to Huang et al. (2023).
1 ., Competing Interests: Declaration of interests The authors declare no competing interests., (Published by Elsevier Inc.)- Published
- 2023
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