Stable Expression of Human H₁‐histamine‐receptor cDNA in Chinese Hamster Ovary Cells

Abstract

A cDNA clone for the histamine HI receptor was isolated from a human lung cDNA library; it encoded a protein of 487 amino acids which showed characteristic features of G-protein-coupled receptors. The percentages of identity of the deduced amino acid sequence with bovine, rat and guinea pig H, histamine receptors were 82.6%, 79.4% and 73.3%, respectively, whereas these percentages decreased to 74.6%, 66% and 56.7% for the amino acid sequence of the third intracellular loop. The human H,-receptor cDNA was transfected into Chinese hamster ovary cells (CHO) via an eukaryotic expression vector ; the receptor protein present on cell membranes specifically bound [3H]mepyramine with a Kd of 3.7 nM. The binding was displaced by H,-histamine-receptor antagonists and histamine. Northern blot analysis indicated the presence of two histamine HI receptor mRNAs of 3.5 kb and 4

International Union of Basic and Clinical Pharmacology. XCVIII. Histamine Receptors

“…The bovine H 1 receptor cDNA was cloned in 1991 by expression cloning in the classic Xenopus oocyte system (Yamashita et al, 1991). This led rapidly to the cloning of the receptor cDNA in several species, including human in 1993(De Backer et al, 1993Moguilevsky et al, 1994). The human gene was subsequently localized to chromosome 3p14-p21 (Le Coniat et al, 1994).…”

Histamine receptors in mammalian retinas

“…The immunogen for the HR1CL antibody (AB5654P; Chemicon) and antiserum (H1R12-S; ADI) was (C)PSFSEIKLRPENPKGDAKK, a peptide (H1R12P; ADI) corresponding to a sequence within the third cytoplasmic loop of the human HR1 (Moguilevsky et al, 1994); cysteine was added for coupling to the carrier protein and was not part of the original sequence. The immunogen for HR3N (H3R32; ADI) was (C)SGALAGDAAAAGGARGFS, a peptide (H3R32P; ADI) corresponding to a sequence within the N-terminus of the human HR3 (Lovenberg et al, 1999(Lovenberg et al, , 2000; cysteine was added for coupling to the carrier protein and was not part of the original sequence.…”

Binding Characteristics of Cetirizine and Levocetirizine to Human H1Histamine Receptors: Contribution of Lys191and Thr194

“…Cloning and stable expression of human histamine H 1 receptors in Chinese hamster ovary (CHO) cells were done in collaboration with Dr. A. Bollen (Department of Applied Genetics, Free University of Brussels, Brussels, Belgium) (Moguilevsky et al, 1994). To perform the mutagenesis, we used the human H 1 receptor cDNA cloned into the plasmid pRc/RSV (pNIV 3604) as template for the synthesis of DNA with site-specific mutations using a polymerase chain reaction strategy.…”

“…Today, four human histamine receptor subtypes have been cloned: H 1 (Moguilevsky et al, 1994), H 2 (Gantz et al, 1991), H 3 (Lovenberg et al, 1999), and, more recently, H 4 (Oda et al, 2000). H 1 receptors induce smooth muscle contraction and increase vascular permeability and H 1 antagonists constitute a medication of choice to alleviate the symptoms of allergies.…”