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. 1977 Jan;59(1):69–81. doi: 10.1172/JCI108623

High affinity binding of 125I-angiotensin II to rat glomerular basement membranes.

J Sraer, L Baud, J P Cosyns, P Verroust, M P Nivez, R Ardaillou
PMCID: PMC333333  PMID: 187623

Abstract

125I-angiotensin II (AII) specifically bound to rat glomerular basement membrane (GBM). The kinetics of binding were similar to those obtained with the total glomeruli. The apparent dissociation constant was close to 50 pM with both preparations. The number of sites related to the amount of protein was two times greater with GBM than with total glomeruli. Since the amount of GBM protein extracted from a given amount of glomerular protein was about 10%, it was possible to estimate the share of the GBM binding sites for AII as representing 20% of the total number present in the entire glomerulus. Binding studies at equilibrium as a function of 125I-AII concentration and competitive binding experiments suggested either multiplicity of the binding sites or cooperativity in the binding reaction. Degradation of 125I-AII in the presence of GBM was slight and did not increase with time. The difference in the degrees of degradation of 125I-AII was too small to account for the observed difference in binding when the results obtained with GBM and isolated glomeruli preparations were compared. 125I-AII binding to GBM was increased after treatment of these membranes with collagenase, slightly diminished with neuraminidase, and almost completely abolished with trypsin suggesting the proteic nature of the receptor. 125I-AII binding to GBM was diminished after incubation of GBM with anti-GBM antibodies as a result of a decrease in the number of binding sites. 125I-AII binding was even more diminished in preparations of glomeruli isolated from rats passively immunized with anti-GBM antibodies when compared with glomeruli from control animals. This resulted from both smaller affinity for AII and decrease in the number of the binding sites. The present data provides evidence for specific binding sites for AII localized on GBM. This is noteworthy since receptors for polypeptide hormones are currently observed on the surface of cell membranes. These findings also suggest a new physiological role for AII which might involve modification of GBM permeability.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Avrameas S. Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies. Immunochemistry. 1969 Jan;6(1):43–52. doi: 10.1016/0019-2791(69)90177-3. [DOI] [PubMed] [Google Scholar]
  2. BENSON J. V., Jr, PATTERSON J. A. ACCELERATED AUTOMATIC CHROMATOGRAPHIC ANALYSIS OF AMINO ACIDS ON A SPHERICAL RESIN. Anal Chem. 1965 Aug;37:1108–1110. doi: 10.1021/ac60228a008. [DOI] [PubMed] [Google Scholar]
  3. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beaufils M., Sraer J., Lepreux C., Ardaillou R. Angiotensin II binding to renal glomeruli from sodium-loaded and sodium-depleted rats. Am J Physiol. 1976 May;230(5):1187–1193. doi: 10.1152/ajplegacy.1976.230.5.1187. [DOI] [PubMed] [Google Scholar]
  5. Blantz R. C., Konnen K. S., Tucker B. J. Angiotensin II effects upon the glomerular microcirculation and ultrafiltration coefficient of the rat. J Clin Invest. 1976 Feb;57(2):419–434. doi: 10.1172/JCI108293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Blau E., Michael A. F. Rat glomerular basement membrane composition and metabolism in aminonucleoside nephrosis. J Lab Clin Med. 1971 Jan;77(1):97–109. [PubMed] [Google Scholar]
  7. Corvol P., Rodbard D., Drouet J., Catt K., Menard J. Monoiodinated angiotensins: preparation and characterization by polyacrylamide gel electrophoresis and isoelectric focusing. Biochim Biophys Acta. 1973 Oct 18;322(2):392–400. doi: 10.1016/0005-2795(73)90314-0. [DOI] [PubMed] [Google Scholar]
  8. Eisenbach G. M., Liew J. B., Boylan J. W., Manz N., Muir P. Effect of angiotensin on the filtration of protein in the rat kidney: a micropuncture study. Kidney Int. 1975 Aug;8(2):80–87. doi: 10.1038/ki.1975.83. [DOI] [PubMed] [Google Scholar]
  9. Flier J. S., Kahn C. R., Roth J., Bar R. S. Antibodies that impair insulin receptor binding in an unusual diabetic syndrome with severe insulin resistance. Science. 1975 Oct 3;190(4209):63–65. doi: 10.1126/science.170678. [DOI] [PubMed] [Google Scholar]
  10. Fong J. S., Drummond K. N. Method for preparation of glomeruli for metabolic studies. J Lab Clin Med. 1968 Jun;71(6):1034–1039. [PubMed] [Google Scholar]
  11. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  12. Hornych H., Beaufils M., Richet G. The effect of exogenous angiotensin on superficial and deep glomeruli in the rat kidney. Kidney Int. 1972 Dec;2(6):336–343. doi: 10.1038/ki.1972.117. [DOI] [PubMed] [Google Scholar]
  13. Jacobs S., Chang K-J, Cuatrecasas P. Estimation of hormone receptor affinity by competitive displacement of labeled ligand: effect of concentration of receptor and of labeled ligand. Biochem Biophys Res Commun. 1975 Sep 16;66(2):687–692. doi: 10.1016/0006-291x(75)90564-1. [DOI] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Osborne M. J., Droz B., Meyer P., Morel F. Angiotensin II: renal localization in glomerular mesangial cells by autoradiography. Kidney Int. 1975 Oct;8(4):245–254. doi: 10.1038/ki.1975.108. [DOI] [PubMed] [Google Scholar]
  16. Patrick J., Lindstrom J. Autoimmune response to acetylcholine receptor. Science. 1973 May 25;180(4088):871–872. doi: 10.1126/science.180.4088.871. [DOI] [PubMed] [Google Scholar]
  17. Sato T., Munakata H., Yoshinaga K., Yosizawa Z. Chemical compositions of glomerular and tubular basement membranes of human kidney. Tohoku J Exp Med. 1975 Apr;115(4):299–306. doi: 10.1620/tjem.115.299. [DOI] [PubMed] [Google Scholar]
  18. Sraer J. D., Sraer J., Ardaillou R., Mimoune O. Evidence for renal glomerular receptors for angiotensin II. Kidney Int. 1974 Oct;6(4):241–246. doi: 10.1038/ki.1974.105. [DOI] [PubMed] [Google Scholar]
  19. Westberg N. G., Michael A. F. Human glomerular basement membrane. Preparation and composition. Biochemistry. 1970 Sep 15;9(19):3837–3846. doi: 10.1021/bi00821a025. [DOI] [PubMed] [Google Scholar]

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