Genomic cloning, heterologous expression and pharmacological characterization of a human histamine H1 receptor.
A human histamine H1 receptor gene lacking introns was isolated by screening a human genomic library with a bovine histamine H1 receptor probe. The deduced protein of 487 amino acids showed characteristic properties of G-protein-coupled receptors. The coding region was subcloned into the expression vector pSVL (Pharmacia), and the resulting construct transfected into COS-7 cells. Binding studies with [3H]pyrilamine on membranes from transfected cells revealed saturable specific binding with a KD of 1.2 nM and a Bmax of 3400 fmol/mg protein. Binding affinities of histamine and known histamine antagonists were similar to those for histamine H1 receptors in guinea-pig cerebellum. In transfected COS-7 cells, histamine induced inositol phosphate formation, that was inhibitable by pyrilamine.
De Backer
M D
MD
Department of Biochemical Pharmacology, Janssen Research Foundation, Beerse, Belgium.
Gommeren
W
W
Moereels
H
H
Nobels
G
G
Van Gompel
P
P
Leysen
J E
JE
Luyten
W H
WH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D015870:Gene Expression; D015698:Genomic Library; D015186:Hominidae / Q000235:genetics*; D006801:Humans; D007438:Introns; D007700:Kinetics; D008969:Molecular Sequence Data; D011738:Pyrilamine / Q000378:metabolism; D051381:Rats; D011969:Receptors, Histamine H1 / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D011969:Receptors, Histamine H1; D011994:Recombinant Proteins; D004247:DNA; D019204:GTP-Binding Proteins; D011738:Pyrilamine