Identification and characterization of a novel angiotensin binding site in cultured vascular smooth muscle cells that is specific for the hexapeptide (3-8) fragment of angiotensin II, angiotensin IV.
This study demonstrates the existence of a previously unrecognized class of angiotensin binding sites on vascular smooth muscle that exhibit high affinity and specificity for the hexapeptide (3-8) fragment of angiotensin II (AngIV). Binding of [125I]AngIV is saturable, reversible and describes a pharmacologic profile that is distinct and separate from the classic AT1 or AT2 angiotensin receptors. Saturation binding studies utilizing cultured vascular smooth muscle cells obtained from bovine aorta (BVSM) revealed that [125I]AngIV bound to a single high affinity site with an associated Hill coefficient of 0.99 +/- 0.003, exhibiting a KD = 1.85 +/- 0.45 nM and a corresponding Bmax = 960 +/- 100 fmol mg-1 protein. Competition binding curves in BVSM demonstrated the following rank order effectiveness: AngIV > AngII(3-7) >> AngIII > Sar1,Ile8 AngII > AngII > AngII(1-7) > AngII(4-8), DuP 753, PD123177. The presence of the non-hydrolyzable GTP analog GTP gamma S, had no effect on [125I]AngIV binding affinity in BVSM. The presence of this novel angiotensin binding site on smooth muscle in high concentration suggests the possibility that this system may play an important, yet unrecognized role in vascular control.
Hall
K L
KL
Department of Veterinary and Comparative Anatomy, Pharmacology, and Physiology, College of Veterinary Medicine, Washington State University, Pullman 99164-6520.
Hanesworth
J M
JM
Ball
A E
AE
Felgenhauer
G P
GP
Hosick
H L
HL
Harding
J W
JW
D000595:Amino Acid Sequence; D000804:Angiotensin II / Q000378:metabolism* / Q000031:analogs & derivatives; D000805:Angiotensin III / Q000031:analogs & derivatives* / Q000378:metabolism; D057911:Angiotensin Receptor Antagonists; D000818:Animals; D001011:Aorta; D001667:Binding, Competitive; D001713:Biphenyl Compounds / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D007093:Imidazoles / Q000378:metabolism; D019808:Losartan; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D011725:Pyridines / Q000378:metabolism; D011869:Radioligand Assay; D011945:Receptors, Angiotensin / Q000378:metabolism*; D013777:Tetrazoles / Q000378:metabolism
D057911:Angiotensin Receptor Antagonists; D001713:Biphenyl Compounds; D007093:Imidazoles; D011725:Pyridines; D011945:Receptors, Angiotensin; D013777:Tetrazoles; D000804:Angiotensin II; C066253:PD 123177; C022929:angiotensin II, 1-des-Asn-2-Arg; D000805:Angiotensin III; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D019808:Losartan