Label-free investigations on the g protein dependent signaling pathways of histamine receptors

Ulla Seibel-Ehlert, Nicole Plank, Asuka Inoue, Guenther Bernhardt, Andrea Strasser

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Abstract: G protein activation represents an early key event in the complex GPCR signal trans-duction process and is usually studied by label-dependent methods targeting specific molecular events. However, the constrained environment of such “invasive” techniques could interfere with biological processes. Although histamine receptors (HRs) represent (evolving) drug targets, their signal transduction is not fully understood. To address this issue, we established a non-invasive dynamic mass redistribution (DMR) assay for the human H1–4Rs expressed in HEK cells, showing excellent signal-to-background ratios above 100 for histamine (HIS) and higher than 24 for inverse agonists with pEC50 values consistent with literature. Taking advantage of the integrative nature of the DMR assay, the involvement of endogenous Gαq/11, Gαs, Gα12/13 and Gβγ proteins was explored, pursuing a two-pronged approach, namely that of classical pharmacology (G protein mod-ulators) and that of molecular biology (Gα knock-out HEK cells). We showed that signal transduction of hH1–4Rs occurred mainly, but not exclusively, via their canonical Gα proteins. For example, in addition to Gαi/o, the Gαq/11 protein was proven to contribute to the DMR response of hH3,4Rs. Moreover, the Gα12/13 was identified to be involved in the hH2R mediated signaling pathway. These results are considered as a basis for future investigations on the (patho)physiological role and the pharmacological potential of H1–4Rs.

Original languageEnglish
Article number9739
JournalInternational journal of molecular sciences
Volume22
Issue number18
DOIs
Publication statusPublished - 2021 Sep

Keywords

  • Dynamic mass redistribution (DMR)
  • G protein coupled receptors (GPCRs)
  • G protein inhibitors
  • G protein knock-out
  • Histamine receptors
  • Label-free
  • Signaling pathways

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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