Molecular Cloning and Functional Characterization of a Mouse Bradykinin B1 Receptor Gene

• Published in: Biochemical and biophysical research communications Vol. 220; no. 1; pp. 219 - 225
• Main Authors: Pesquero, João B., Pesquero, Jorge L., Oliveira, Suzana M., Roscher, Adelbert A., Metzger, Rainer, Ganten, Detlev, Bader, Michael
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 07-03-1996
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cloning, Molecular; DNA Primers - genetics; Humans; Kinetics; Mice; Molecular Sequence Data; Rabbits; Receptor, Bradykinin B1; Receptors, Bradykinin - drug effects; Receptors, Bradykinin - genetics; Receptors, Bradykinin - metabolism; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sequence Homology, Amino Acid; Tissue Distribution
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1996.0384

The gene encoding a putative mouse bradykinin B1 receptor was cloned from a genomic library by low stringency screening. Analysis of two isolated clones revealed a region which contains an open reading frame uninterrupted by introns and encodes a 334 amino acid protein, which exhibits seven potential transmembrane domains and is 68% identical to the human and rabbit bradykinin B1 receptors. Lipopolysaccharide-treatment induces B1 receptor transcripts in the heart, liver and lung. Stable expression of the coding region in COS-7 cells resulted in high levels of binding sites for the specific B1 ligand des-Arg10kallidin (Kd= 1.3 nM; Bmax= 51 fmol/mg protein). The rank order of affinity of the receptor for the agonists and antagonists was: des-Arg9BK>des-Arg9Leu8BK>des-Arg10kallidin⪢Hoe-140=bradykinin. Functional coupling of the cloned receptor was demonstrated by the dose-dependent effect of des-Arg9BK on the extracellular acidification rate in stably transfected COS-7 cells. This effect was not produced by bradykinin and could be blocked by the B1 antagonist des-Arg9Leu8BK.