Publication details for Dr P Chazot

Van Rijn, RM., Chazot, PL., Shenton, FC., Sansuk, K., Bakker, RA. & Leurs, R. (2006). Oligomerization of recombinant and endogenously expressed human histamine H4 receptors. Molecular Pharmacology 70(2): 604-615.

• Publication type: Journal Article
• ISSN/ISBN: 0026-895X, 1521-0111
• DOI: 10.1124/mol.105.020818
• Further publication details on publisher web site
• Durham Research Online (DRO) - may include full text

Author(s) from Durham

• Dr Fiona Cameron Shenton
• Dr P Chazot

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.

Notes

Joint first authors