Oligomerization of recombinant and endogenously expressed human histamine H4 receptors

Van Rijn, RM.; Chazot, PL.; Shenton, FC.; Sansuk, K.; Bakker, RA.; Leurs, R.

Abstract

In this study, we report the homo- and hetero-oligomerization of the human histamine H4R by both biochemical (Western blot and immobilized metal affinity chromatography) and biophysical [bioluminescence resonance energy transfer and time-resolved fluorescence resonance energy transfer (tr-FRET)] techniques. The H4R receptor is the most recently discovered member of the histamine family of G-protein-coupled receptors. Using specific polyclonal antibodies raised against the C-terminal tail of the H4R, we demonstrate the presence of H4R oligomers in human embryonic kidney 293 and COS-7 cells heterologously overexpressing H4Rs and putative native H4R oligomers in human phytohaemagglutinin blasts endogenously expressing H4Rs. Moreover, we show that H4R homo-oligomers are formed constitutively, are formed at low receptor densities (300 fmol/mg of protein), and are present at the cell surface, as detected by tr-FRET. The formation of these oligomers is independent of N-glycosylation and is not modulated by H4R ligands, covering the full spectrum of agonists, neutral antagonists, and inverse agonists. Although we show H4R homo-oligomer formation at physiological expression levels, the detection of H1R-H4R hetero-oligomers was achieved only at higher H1R expression levels and are most likely not physiologically relevant.

Citation

Van Rijn, R., Chazot, P., Shenton, F., Sansuk, K., Bakker, R., & Leurs, R. (2006). Oligomerization of recombinant and endogenously expressed human histamine H4 receptors. Molecular Pharmacology, 70(2), 604-615. https://doi.org/10.1124/mol.105.020818

• Journal Article Type: Article
• Publication Date: Apr 1, 2006
• Deposit Date: May 21, 2007
• Journal: Molecular Pharmacology
• Print ISSN: 0026-895X
• Electronic ISSN: 1521-0111
• Publisher: American Society for Pharmacology and Experimental Therapeutics (ASPET)
• Peer Reviewed: Not Peer Reviewed
• Volume: 70
• Issue: 2
• Pages: 604-615
• DOI: https://doi.org/10.1124/mol.105.020818
• Public URL: https://durham-repository.worktribe.com/output/1589849