Pharmacological and functional characterisation of the wild-type and site-directed mutants of the human H1 histamine receptor stably expressed in CHO cells

J Recept Signal Transduct Res. 1995 Jan-Mar;15(1-4):91-102. doi: 10.3109/10799899509045210.

Abstract

A cDNA clone for the human histamine H1 receptor was isolated from a lung cDNA library and stably expressed in CHO cells. The recombinant receptor protein present in the cell membranes, displayed the functional and binding characteristics of histamine H1 receptors. Mutation of Ser155 to Ala in the fourth transmembrane domain did not significantly change the affinity of the receptor for histamine and H1 antagonists. However, mutation of the fifth transmembrane Asn198 to Ala resulted in a dramatic decrease of the affinity for histamine binding, and for the histamine-induced polyphosphoinositides breakdown, whereas the affinity towards antagonists was not significantly modified. In addition, mutation of another fifth transmembrane amino acid, Thr194 to Ala also diminished, but to a lesser extent, the affinity for histamine. These data led us to propose a molecular model for histamine interaction with the human H1 receptor. In this model, the amide moiety of Asn198 and the hydroxyl group of Thr194 are involved in hydrogen bonding with the nitrogen atoms of the imidazole ring of histamine. Moreover, mutation of Thr194 to Ala demonstrated that this residue is responsible for the discrimination between enantiomers of cetirizine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites / genetics
  • CHO Cells
  • Cloning, Molecular
  • Cricetinae
  • Histamine / metabolism
  • Histamine / pharmacology
  • Humans
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Phosphatidylinositol Phosphates / metabolism
  • Pyrilamine / metabolism
  • Receptors, Histamine H1 / drug effects
  • Receptors, Histamine H1 / genetics*
  • Receptors, Histamine H1 / metabolism*
  • Transfection

Substances

  • Phosphatidylinositol Phosphates
  • Receptors, Histamine H1
  • Histamine
  • Pyrilamine