GPCR kinase knockout cells reveal the impact of individual GRKs on arrestin binding and GPCR regulation

Nat Commun. 2022 Jan 27;13(1):540. doi: 10.1038/s41467-022-28152-8.


G protein-coupled receptors (GPCRs) activate G proteins and undergo a complex regulation by interaction with GPCR kinases (GRKs) and the formation of receptor-arrestin complexes. However, the impact of individual GRKs on arrestin binding is not clear. We report the creation of eleven combinatorial HEK293 knockout cell clones lacking GRK2/3/5/6, including single, double, triple and the quadruple GRK knockout. Analysis of β-arrestin1/2 interactions for twelve GPCRs in our GRK knockout cells enables the differentiation of two main receptor subsets: GRK2/3-regulated and GRK2/3/5/6-regulated receptors. Furthermore, we identify GPCRs that interact with β-arrestins via the overexpression of specific GRKs even in the absence of agonists. Finally, using GRK knockout cells, PKC inhibitors and β-arrestin mutants, we present evidence for differential receptor-β-arrestin1/2 complex configurations mediated by selective engagement of kinases. We anticipate our GRK knockout platform to facilitate the elucidation of previously unappreciated details of GRK-specific GPCR regulation and β-arrestin complex formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arrestin / metabolism*
  • G-Protein-Coupled Receptor Kinases / metabolism*
  • GTP-Binding Proteins / metabolism
  • HEK293 Cells
  • Humans
  • Phosphorylation
  • Receptors, G-Protein-Coupled / metabolism*
  • Signal Transduction / physiology
  • beta-Arrestin 1 / metabolism
  • beta-Arrestin 2 / metabolism


  • Arrestin
  • Receptors, G-Protein-Coupled
  • beta-Arrestin 1
  • beta-Arrestin 2
  • G-Protein-Coupled Receptor Kinases
  • GTP-Binding Proteins