Open AccessStable Expression of Human H 1 ‐histamine‐receptor cDNA in Chinese Hamster Ovary Cells
Author(s) -
Moguilevsky Nicole,
Varsalona Francesca,
Noyer Michel,
Gillard Michel,
Guillaume JanPaul,
Garcia Lida,
Szpirer Claude,
Szpirer Josiane,
Bollen Alex
Publication year - 1994
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1994.00489.x
Subject(s) - microbiology and biotechnology , chinese hamster ovary cell , complementary dna , biology , histamine receptor , histamine h1 receptor , receptor , histamine , cdna library , northern blot , histamine h4 receptor , histamine h2 receptor , gene , biochemistry , endocrinology , antagonist
A cDNA clone for the histamine H 1 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 1 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 1 ‐receptor cDNA was transfected into Chinese hamster ovary cells (CHO) via an eukaryotic expression vector; the receptor protein present on cell membranes specifically bound [ 3 H]mepyramine with a K d of 3.7 nM. The binding was displaced by H 1 ‐histamine‐receptor antagonists and histamine. Northern blot analysis indicated the presence of two histamine H 1 receptor mRNAs of 3.5 kb and 4.1 kb in various human tissues and an additional mRNA of 4.8 kb restricted to the human brain. Finally, by means of somatic cell hybrids segregating either human or rat chromosomes, the gene for histamine H 1 receptor was found to reside on human chromosome 3 and rat chromosome 4.