Cloning and pharmacological characterization of a human bradykinin (BK-2) receptor.
A human BK-2 bradykinin receptor was cloned from the lung fibroblast cell line CCD-16Lu. The cDNA clone encodes a 364 amino acid protein that has the characteristics of a seven transmembrane domain G-protein coupled receptor. The predicted amino acid sequence of the human BK-2 receptor is 81% identical to the smooth muscle rat BK-2 receptor (1). Transfection of the human BK-2 receptor cDNA into COS-7 cells results in the expression of high levels of specific BK binding sites. Saturation binding analysis indicates that the human BK-2 receptor expressed in COS-7 cells binds BK with a KD of 0.13 nM. Pharmacological characterization of the expressed BK receptor is consistent with the cDNA encoding a receptor of the BK-2 subtype. The BK-2 receptor antagonist Hoe 140 (2), D-Arg0[Hyp3, Thi5, D-Tic7, Oic8]BK has a high affinity (IC50 = 65 pM) for the cloned human receptor. The tissue distribution of the human BK-2 receptor was analyzed by competitive PCR with human tissue cDNA and is similar to that determined for the BK-2 receptor in the rat.
Hess
J F
JF
Merck Research Laboratories, Department of Molecular Pharmacology and Biochemistry, Rahway, N.J. 07065.
Borkowski
J A
JA
Young
G S
GS
Strader
C D
CD
Ransom
R W
RW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D001920:Bradykinin / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D006031:Glycosylation; D006801:Humans; D007700:Kinetics; D008168:Lung; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D018002:Receptors, Bradykinin; D017981:Receptors, Neurotransmitter / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D018002:Receptors, Bradykinin; D017981:Receptors, Neurotransmitter; D011494:Protein Kinases; D001920:Bradykinin