Phosphorylation barcodes direct biased chemokine signaling at CXCR3

Dylan S. Eiger, Jeffrey S. Smith, Tujin Shi, Tomasz Maciej Stepniewski, Chia Feng Tsai, Christopher Honeycutt, Noelia Boldizsar, Julia Gardner, Carrie D. Nicora, Ahmed M. Moghieb, Kouki Kawakami, Issac Choi, Chloe Hicks, Kevin Zheng, Anmol Warman, Priya Alagesan, Nicole M. Knape, Ouwen Huang, Justin D. Silverman, Richard D. SmithAsuka Inoue, Jana Selent, Jon M. Jacobs, Sudarshan Rajagopal

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

G protein-coupled receptor (GPCR)-biased agonism, selective activation of certain signaling pathways relative to others, is thought to be directed by differential GPCR phosphorylation “barcodes.” At chemokine receptors, endogenous chemokines can act as “biased agonists”, which may contribute to the limited success when pharmacologically targeting these receptors. Here, mass spectrometry-based global phosphoproteomics revealed that CXCR3 chemokines generate different phosphorylation barcodes associated with differential transducer activation. Chemokine stimulation resulted in distinct changes throughout the kinome in global phosphoproteomics studies. Mutation of CXCR3 phosphosites altered β-arrestin 2 conformation in cellular assays and was consistent with conformational changes observed in molecular dynamics simulations. T cells expressing phosphorylation-deficient CXCR3 mutants resulted in agonist- and receptor-specific chemotactic profiles. Our results demonstrate that CXCR3 chemokines are non-redundant and act as biased agonists through differential encoding of phosphorylation barcodes, leading to distinct physiological processes.

Original languageEnglish (US)
Pages (from-to)362-382.e8
JournalCell Chemical Biology
Volume30
Issue number4
DOIs
StatePublished - Apr 20 2023

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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