Molecular cloning and expression of rat bradykinin B1 receptor.
The gene encoding rat bradykinin B1 receptor has been cloned by using a partial rat B1 cDNA probe. The rat B1 receptor gene contains two exons and the entire coding region is within the second exon. The 5'-flanking region of the rat B1 receptor gene contains several putative transcriptional regulatory sites including TATA box, cAMP response element, NF-kappaB and AP-1. It showed promoter activity inducible by lipopolysaccharide in vascular smooth muscle cells. Rat B1 receptor mRNA was found to be alternatively spliced and induced by lipopolysaccharide treatment in a wide range of tissues, such as the salivary gland, testis, kidney, lung, heart, prostate and aorta. The deduced rat B1 receptor amino acid sequence is 71% homologous to human and rabbit counterparts, and 89% homologous to the mouse counterpart. The expressed B1 receptor in HEK293 cells displayed a rank order of affinity for the kinin peptides: des-Arg9-BK>Lys-des-Arg9-BK approximately des-Arg9, Leu8-BK>Sar-Tyr-epsilonAhx-Lys-[D-betaNal7, Ile8]-des-Arg9-BK>Sar-Tyr-epsilonAhx-Lys-des-Arg9-BK>>BK>> Hoe140. These results indicate that the cloned gene encodes a functional rat B1 receptor.
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Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC 29425, USA.
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D006801:Humans; D007700:Kinetics; D007705:Kinins / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011817:Rabbits; D051381:Rats / Q000235:genetics*; D043783:Receptor, Bradykinin B1; D018002:Receptors, Bradykinin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D007705:Kinins; D043783:Receptor, Bradykinin B1; D018002:Receptors, Bradykinin; D011993:Recombinant Fusion Proteins