Characterization of angiotensin II binding sites in the human term placenta.
Specific angiotensin II (AII) binding sites were identified and characterized in membranes from human term placenta. The binding of iodinated [125I](Sar1)AII was time-dependent and saturable; it could be totally reversed on addition of unlabelled (Sar1)AII and GTP + NaCl. Scatchard plot analysis of dose-dependent [125I](Sar1)AII binding indicated the presence of a single class of binding sites with an equilibrium dissociation constant of 0.27 +/- 0.06 nM and a maximum binding capacity of 38.4 +/- 4.3 fmol/mg protein. The affinity of five AII analogues for the placental receptor was determined in competitive binding assays; the order of inhibitory potency was: (Sar1)AII greater than (Sar1, Ile8)AII approximately (Sar1, Ala8)AII approximately AII greater than angiotensin I greater than (Des-Phe8)AII. (Sar1)AII did not cause any significant change in the basal or stimulated adenylate cyclase activity. In order to investigate the subunit molecular structure of the placenta AII receptor, membranes were covalently labelled with the photoaffinity ligand [125I](Sar1, (4N3Phe)8)AII. Sodium dodecylsulfate-polyacrylamide gel electrophoresis followed by autoradiography showed that the labelling was specifically incorporated into a protein of Mr 92,000 in the presence or absence of dithiothreitol. It therefore appears that the AII receptor from human placenta has the same binding and pharmacological properties as other well-known AII receptors; by contrast, it is characterized by a significantly higher molecular weight, pointing out that structural differences in AII receptors may exist between species.
Tencé
M
M
Centre CNRS-INSERM de Pharmacologie-Endocrinologie, Montpellier, France.
Petit
A
A
D000262:Adenylyl Cyclases / Q000378:metabolism; D000804:Angiotensin II / Q000032:analysis / Q000378:metabolism*; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D010920:Placenta / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D011247:Pregnancy
D000804:Angiotensin II; D000262:Adenylyl Cyclases