Perturbation of the interactions of calmodulin with GRK5 using a natural product chemical probe

Tyler S. Beyett, Amy E. Fraley, Emily Labudde, Dhabaleswar Patra, Ryan C. Coleman, Akito Eguchi, Alisa Glukhova, Qiuyan Chen, Robert M. Williams, Walter J. Koch, David H. Sherman, John J.G. Tesmer

Research output: Contribution to journalArticleResearchpeer-review

4 Citations (Scopus)

Abstract

G protein-coupled receptor (GPCR) kinases (GRKs) are responsible for initiating desensitization of activated GPCRs. GRK5 is potently inhibited by the calcium-sensing protein calmodulin (CaM), which leads to nuclear translocation of GRK5 and promotion of cardiac hypertrophy. Herein, we report the architecture of the Ca2+·CaM–GRK5 complex determined by small-angle X-ray scattering and negative-stain electron microscopy. Ca2+·CaM binds primarily to the small lobe of the kinase domain of GRK5 near elements critical for receptor interaction and membrane association, thereby inhibiting receptor phosphorylation while activating the kinase for phosphorylation of soluble substrates. To define the role of each lobe of Ca2+·CaM, we utilized the natural product malbrancheamide as a chemical probe to show that the C-terminal lobe of Ca2+·CaM regulates membrane binding while the N-terminal lobe regulates receptor phosphorylation and kinase domain activation. In cells, malbrancheamide attenuated GRK5 nuclear translocation and effectively blocked the hypertrophic response, demonstrating the utility of this natural product and its derivatives in probing Ca2+·CaM-dependent hypertrophy.

Original languageEnglish
Pages (from-to)15895-15900
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number32
DOIs
Publication statusPublished - 6 Aug 2019
Externally publishedYes

Keywords

  • Calmodulin
  • G protein-coupled receptor kinase 5
  • Hypertrophy
  • Malbrancheamide

Cite this