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. 1991 Oct;104(2):321–326. doi: 10.1111/j.1476-5381.1991.tb12429.x

Neuropeptide Y (NPY) metabolism by endopeptidase-2 hinders characterization of NPY receptors in rat kidney.

J S Price 1, A J Kenny 1, N S Huskisson 1, M J Brown 1
PMCID: PMC1908564  PMID: 1665730

Abstract

1. Despite the observation of pharmacological responses to neuropeptide Y (NPY) in mammalian kidneys, there are species differences in the ease with which specific NPY binding sites can be demonstrated; we have investigated whether this can be explained by differential metabolism of NPY by a membrane-bound peptidase. 2. NPY receptors were identified on cell membranes isolated from the rabbit kidney (KD = 97 +/- 16 pM, Bmax = 290 +/- 30 fmol mg-1 protein), and this preparation did not degrade [125I]-NPY. However, a similar preparation of cell membranes from the rat kidney exhibited a much lower apparent receptor affinity (IC50 approximately 30 nM); these membranes rapidly degraded [125I]-NPY to fragments which did not bind NPY receptors in either tissue. 3. [125I]-NPY binding sites were revealed in the rat kidney when degradation was inhibited by insulin B chain. Chelating agents also inhibited degradation, but interfered with receptor binding. Binding sites could not be demonstrated in sections of rat kidney, even in the presence of insulin B chain. 4. The difference in degradative activity between rat and rabbit renal cell membranes, inhibition of degradation by chelating agents and insulin B chain, and insensitivity to phosphoramidon suggest that the enzyme responsible was endopeptidase-2, and this was confirmed by comparing the hydrolysis of [125I]-NPY by purified enzyme with rat renal tissue. Activity of this enzyme explains the difficulties encountered demonstrating receptors in the rat kidney. 5. Renal cell membranes from the mouse digested [125I]-NPY in a similar manner and this may be due to the closely related enzyme, meprin.(ABSTRACT TRUNCATED AT 250 WORDS)

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Selected References

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  1. Allen J. M., Raine A. E., Ledingham J. G., Bloom S. R. Neuropeptide Y: a novel renal peptide with vasoconstrictor and natriuretic activity. Clin Sci (Lond) 1985 Apr;68(4):373–377. doi: 10.1042/cs0680373. [DOI] [PubMed] [Google Scholar]
  2. Ballesta J., Polak J. M., Allen J. M., Bloom S. R. The nerves of the juxtaglomerular apparatus of man and other mammals contain the potent peptide NPY. Histochemistry. 1984;80(5):483–485. doi: 10.1007/BF00495438. [DOI] [PubMed] [Google Scholar]
  3. Barnes K., Ingram J., Kenny A. J. Proteins of the kidney microvillar membrane. Structural and immunochemical properties of rat endopeptidase-2 and its immunohistochemical localization in tissues of rat and mouse. Biochem J. 1989 Dec 1;264(2):335–346. doi: 10.1042/bj2640335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beynon R. J., Shannon J. D., Bond J. S. Purification and characterization of a metallo-endoproteinase from mouse kidney. Biochem J. 1981 Dec 1;199(3):591–598. doi: 10.1042/bj1990591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DeBlasi A., O'Reilly K., Motulsky H. J. Calculating receptor number from binding experiments using same compound as radioligand and competitor. Trends Pharmacol Sci. 1989 Jun;10(6):227–229. doi: 10.1016/0165-6147(89)90266-6. [DOI] [PubMed] [Google Scholar]
  6. Edvinsson L., Ekblad E., Håkanson R., Wahlestedt C. Neuropeptide Y potentiates the effect of various vasoconstrictor agents on rabbit blood vessels. Br J Pharmacol. 1984 Oct;83(2):519–525. doi: 10.1111/j.1476-5381.1984.tb16516.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ekblad E., Edvinsson L., Wahlestedt C., Uddman R., Håkanson R., Sundler F. Neuropeptide Y co-exists and co-operates with noradrenaline in perivascular nerve fibers. Regul Pept. 1984 Apr;8(3):225–235. doi: 10.1016/0167-0115(84)90064-8. [DOI] [PubMed] [Google Scholar]
  8. Evéquoz D., Waeber B., Corder R., Nussberger J., Gaillard R., Brunner H. R. Markedly reduced blood pressure responsiveness in endotoxemic rats; reversal by neuropeptide Y. Life Sci. 1987 Dec 7;41(23):2573–2580. doi: 10.1016/0024-3205(87)90442-5. [DOI] [PubMed] [Google Scholar]
  9. Kassis S., Olasmaa M., Terenius L., Fishman P. H. Neuropeptide Y inhibits cardiac adenylate cyclase through a pertussis toxin-sensitive G protein. J Biol Chem. 1987 Mar 15;262(8):3429–3431. [PubMed] [Google Scholar]
  10. Kenny A. J., Fulcher I. S., Ridgwell K., Ingram J. Microvillar membrane neutral endopeptidases. Acta Biol Med Ger. 1981;40(10-11):1465–1471. [PubMed] [Google Scholar]
  11. Kenny A. J., Ingram J. Proteins of the kidney microvillar membrane. Purification and properties of the phosphoramidon-insensitive endopeptidase ('endopeptidase-2') from rat kidney. Biochem J. 1987 Jul 15;245(2):515–524. doi: 10.1042/bj2450515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Leys K., Schachter M., Sever P. Autoradiographic localisation of NPY receptors in rabbit kidney: comparison with rat, guinea-pig and human. Eur J Pharmacol. 1987 Feb 10;134(2):233–237. doi: 10.1016/0014-2999(87)90171-3. [DOI] [PubMed] [Google Scholar]
  13. Lundberg J. M., Hemsén A., Larsson O., Rudehill A., Saria A., Fredholm B. B. Neuropeptide Y receptor in pig spleen: binding characteristics, reduction of cyclic AMP formation and calcium antagonist inhibition of vasoconstriction. Eur J Pharmacol. 1988 Jan 5;145(1):21–29. doi: 10.1016/0014-2999(88)90344-5. [DOI] [PubMed] [Google Scholar]
  14. Lundberg J. M., Stjarne L. Neuropeptide Y (NPY) depresses the secretion of 3H-noradrenaline and the contractile response evoked by field stimulation, in rat vas deferens. Acta Physiol Scand. 1984 Mar;120(3):477–479. doi: 10.1111/j.1748-1716.1984.tb07410.x. [DOI] [PubMed] [Google Scholar]
  15. Lundberg J. M., Terenius L., Hökfelt T., Martling C. R., Tatemoto K., Mutt V., Polak J., Bloom S., Goldstein M. Neuropeptide Y (NPY)-like immunoreactivity in peripheral noradrenergic neurons and effects of NPY on sympathetic function. Acta Physiol Scand. 1982 Dec;116(4):477–480. doi: 10.1111/j.1748-1716.1982.tb07171.x. [DOI] [PubMed] [Google Scholar]
  16. Pernow J., Lundberg J. M., Kaijser L. Vasoconstrictor effects in vivo and plasma disappearance rate of neuropeptide Y in man. Life Sci. 1987 Jan 5;40(1):47–54. doi: 10.1016/0024-3205(87)90251-7. [DOI] [PubMed] [Google Scholar]
  17. Pernow J., Svenberg T., Lundberg J. M. Actions of calcium antagonists on pre- and postjunctional effects of neuropeptide Y on human peripheral blood vessels in vitro. Eur J Pharmacol. 1987 Apr 14;136(2):207–218. doi: 10.1016/0014-2999(87)90712-6. [DOI] [PubMed] [Google Scholar]
  18. Reynolds E. E., Yokota S. Neuropeptide Y receptor-effector coupling mechanisms in cultured vascular smooth muscle cells. Biochem Biophys Res Commun. 1988 Mar 15;151(2):919–925. doi: 10.1016/s0006-291x(88)80369-3. [DOI] [PubMed] [Google Scholar]
  19. Sheikh S. P., Håkanson R., Schwartz T. W. Y1 and Y2 receptors for neuropeptide Y. FEBS Lett. 1989 Mar 13;245(1-2):209–214. doi: 10.1016/0014-5793(89)80223-6. [DOI] [PubMed] [Google Scholar]
  20. Stephenson S. L., Kenny A. J. Metabolism of neuropeptides. Hydrolysis of the angiotensins, bradykinin, substance P and oxytocin by pig kidney microvillar membranes. Biochem J. 1987 Jan 1;241(1):237–247. doi: 10.1042/bj2410237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stephenson S. L., Kenny A. J. The metabolism of neuropeptides. Hydrolysis of peptides by the phosphoramidon-insensitive rat kidney enzyme 'endopeptidase-2' and by rat microvillar membranes. Biochem J. 1988 Oct 1;255(1):45–51. doi: 10.1042/bj2550045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sterchi E. E., Green J. R., Lentze M. J. Non-pancreatic hydrolysis of N-benzoyl-l-tyrosyl-p-aminobenzoic acid (PABA-peptide) in the human small intestine. Clin Sci (Lond) 1982 May;62(5):557–560. doi: 10.1042/cs0620557. [DOI] [PubMed] [Google Scholar]
  23. Stjärne L., Lundberg J. M., Astrand P. Neuropeptide Y--a cotransmitter with noradrenaline and adenosine 5'-triphosphate in the sympathetic nerves of the mouse vas deferens? A biochemical, physiological and electropharmacological study. Neuroscience. 1986 May;18(1):151–166. doi: 10.1016/0306-4522(86)90184-3. [DOI] [PubMed] [Google Scholar]
  24. Tatemoto K. Neuropeptide Y: complete amino acid sequence of the brain peptide. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5485–5489. doi: 10.1073/pnas.79.18.5485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Undén A., Tatemoto K., Mutt V., Bartfai T. Neuropeptide Y receptor in the rat brain. Eur J Biochem. 1984 Dec 17;145(3):525–530. doi: 10.1111/j.1432-1033.1984.tb08588.x. [DOI] [PubMed] [Google Scholar]
  26. Wahlestedt C., Yanaihara N., Håkanson R. Evidence for different pre-and post-junctional receptors for neuropeptide Y and related peptides. Regul Pept. 1986 Feb;13(3-4):307–318. doi: 10.1016/0167-0115(86)90048-0. [DOI] [PubMed] [Google Scholar]

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