Klinische Wochenschrift

, Volume 66, Issue 17, pp 752–759 | Cite as

Isolation and structural analysis of “Urodilatin”, a new peptide of the cardiodilatin-(ANP)-family, extracted from human urine

  • P. Schulz-Knappe
  • K. Forssmann
  • F. Herbst
  • D. Hock
  • R. Pipkorn
  • W. G. Forssmann


Two major forms of cardiac peptides have been established in the last few years: (a) a prohormone of 126 amino acids (CDD/ANP-1-126) in the endocrine heart and (b) the circulating CDD/ANP-99-126 (=alpha ANP) in blood plasma. The method we applied earlier to isolate the circulating form of cardiodilatin from human blood was used to detect and analyze the biologically active, predominant form of the same polypeptide family excreted by the kidneys. Each step of the isolation procedure was followed up by a bioassay using an in vitro vascular smooth muscle relaxation test and a highly specific RIA against cardiodilatin (CDD-99-126) for the initial purification steps. The polypeptides excreted in 1000 1 of normal human urine were adsorbed to 2.5 kg of alginic acid, and after elution and lyophilization processed on a G-25 Sephadex column. The obtained crude polypeptide fractions were applied to ion-exchange chromatography. Thereafter four steps of HPLC were carried out to purify the polypeptide which was the suggested form of cardiodilatin (CDD) in human urine. The amino acid analysis and gas phase sequence analysis showed that the main form of urinary cardiodilatin is a 32 amino acid residue containing molecule, cardiodilatin-95–126. The molecule is N-terminally extended compared to the circulating CDD-99-126. This suggests that the analyzed urinary peptide is not the residual plasma form, filtrated and renally cleared from blood, but probably a polypeptide produced and processed in the kidney tubules and cleaved by a different postranslational process. Therefore, this vasorelaxant polypeptide is called urodilatin.

Key words

Cardiodilation (CDD) Urodilatin Posttranslational Processing Cardiac Hormones Atrial Natriuretic Polypeptide (ANP) Urine Polypeptide Hormone Amino Acid Sequence 



atrial natriuretic polypeptide




high performance liquid chromatography




messenger ribonucleic acid




optical density




reversed phase


trifluoro-acetic acid


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Bindlingmeyer BA, Cohen SA, Tarvin TL (1984) Rapid analysis of amino acids using pre-column derivatization. J Chromatogr 336:93–104Google Scholar
  2. 2.
    Cantin M, Genest J (1985) The heart and the atrial natriuretic factor. Endocr Rev 6:107–127Google Scholar
  3. 3.
    Feller S, Rippegather G, Forssmann WG (1988) Expression of the CDD/ANP gene in human and porcine kidney tubules. (in preparation)Google Scholar
  4. 4.
    Feller S, Meyer M, Hock D, Forssmann WG (1988) Extraauriculäre Lokalisation von Cardiodilatin. Acta anat (in press)Google Scholar
  5. 5.
    Flügge G, Inagami T, Fuchs E (1987) Atrial natriuretic peptide detected by immunocytochemistry in peripheral organs of Tupaia belangeri. Histochemistry 86:479–483Google Scholar
  6. 6.
    Flynn TG, De Bold ML, De Bold AJ (1983) The amino acid sequence of an atrial peptide with potent diuretic and natriuretic properties. Biochem Biophys Res Comm 117:859–865Google Scholar
  7. 7.
    Forssmann K, Hock D, Herbst F, Schulz-Knappe P, Talartschik J, Scheler F, Forssmann WG (1986) Isolation and structural analysis of the circulating human cardiodilatin (Alpha ANP). Klin Wochenschr 64:1276–1280Google Scholar
  8. 8.
    Forssmann WG (1986) Cardiac hormones. I. Review on the morphology, biochemistry, and molecular biology of the endocrine heart. Eur J Clin Invest 16:439–451Google Scholar
  9. 9.
    Forssmann WG, Birr C, Carlquist M, Christmann M, Finke R, Henschen A, Hock D, Kirchheim H, Kreye V, Lottspeich F, Metz J, Mutt V, Reinecke M (1984) The auricular myocardiocytes of the heart constitute an endocrine organ. Characterization of a porcine cardiac peptide hormone, cardiodilatin-126. Cell Tissue Res 238:425–430Google Scholar
  10. 10.
    Forssmann WG, Hock D, Lottspeich F, Henschen A, Kreye V, Christmann M, Reinecke M, Metz J, Carlquist M, Mutt V (1983) The right auricle of the heart is an endocrine organ. Cardiodilatin as a peptide hormone candidate. Anat Embryol 168:307–313Google Scholar
  11. 11.
    Forssmann WG, Hock D, Mutt V (1986) Cardiac hormones: morphology and biochemistry. Klin Wochenschr 64(Suppl VI):4–12Google Scholar
  12. 12.
    Gagelmann M, Hock D, Forssmann WG (1988) Urodilatin (CDD/ANP-95-126) is not biologically inactivated by a peptidase from dog kidney cortex membranes in contrast to atrial natriuretic peptide/cardiodilatin (α-hANP/CDD-99-126). FEBS letters 233:249–254Google Scholar
  13. 13.
    Greenberg BD, Bencen GH, Seilhamer JJ, Lewicki JA, Fiddes JC (1984) Nucleotide sequence of the gene encoding human atrial natriuretic factor precursor. Nature 312:656–658Google Scholar
  14. 14.
    Greenwald J, McLaughlin L, Needleman P (1986) Atriopeptide excretion in human urine. Fed Proc 45:912Google Scholar
  15. 15.
    Hock D, Schriek U, Fey E, Forssmann WG, Mutt V (1987) Isolation of bovine cardiodilatin by fast protein liquid chromatography and reversed-phase high-performance liquid chromatography. J Chromatography 397:347–353Google Scholar
  16. 16.
    Kangawa K, Matsuo H (1984) Purification and complete amino acid sequence of alpha-human atrial natriuretic polypeptide (alpha-hANP). Biochem Biophys Res Comm 118:131–139Google Scholar
  17. 17.
    Koehn JA, Norman JA, Jones BN, LeSueur L, Sakane Y, Ghai RD (1987) Degradation of atrial natriuretic factor by kidney cortex membranes. J Biol Chem 262:11623–11627Google Scholar
  18. 18.
    Lottspeich F (1981) Identification of phenylthiohydantoin amino acids for microscale sequencing. In: Lottspeich F, Henschen A, Hupe KP (eds) High performance liquid chromatography in protein and peptide chemistry. Walter de Gruyter, Berlin New York, pp 259–268Google Scholar
  19. 19.
    Marumo F, Sakamoto H, Ando K, Ishigami T, Kawakami M (1986) A highly sensitive radioimmunoassay of atrial natriuretic peptide (ANP) in human plasma and urine. Biochem Biophys Res Commun 137:231–236Google Scholar
  20. 20.
    Marumo F, Umetani N, Sakamoto H, Ando K, Ishigami T (1987) Characteristics of atrial natriuretic peptide (ANP) in plasma and urine in renal diseases. Kidney Internat 31:278Google Scholar
  21. 21.
    Mutt V (1978) Hormone isolation. In: Bloom SR (ed) Gut hormones. Churchill Livingstone, Edinburgh London New York, pp 21–27Google Scholar
  22. 22.
    Olins GM, Spear KL, Siegel NR, Zurcher-Neely HA (1987) Inactivation of atrial natriuretic factor by the renal brush border. Biochim Biophys Acta 901:97–100Google Scholar
  23. 23.
    Rippegather G (1988) Pathomorphology of the endocrine heart. In: Forssmann WG, Scheuermann DW (ed) Functional morphology of the endocrine heart. (in press)Google Scholar
  24. 24.
    Sakamoto M, Nakao K, Kihara M, Morii N, Sugawara A, Suda M, Shimokura M, Kiso Y, Yamori Y, Imura H (1985) Existence of atrial natriuretic polypeptide in kidney. Biochem Biophys Res Commun 128:1281–1287Google Scholar
  25. 25.
    Stephenson SL, Kenny AJ (1987) The hydrolysis of a-human atrial natriuretic peptide by pig kidney microvillar membranes is initiated by endopeptidase-24.11. Biochem J 243:183–187Google Scholar
  26. 26.
    Strickler JE, Hunkapiller MW, Wilson KJ (1984) Utility of the gas-phase sequencer for both liquid- and solid-phase degradation of proteins and peptides at low picomole levels. Anal Biochem 140:553–566Google Scholar
  27. 27.
    Weil J, Heim JM, Döhlemann C, Bühlmeyer K, Gerzer R (1987) Konzentration von atrialem natriuretischem Peptid and cGMP im Plasma und Urin bei herzkranken Kindern. In: Kreye VAW, Bussmann WD (ed) ANP — Atriales natriuretisches Peptid und das kardiovaskuläre System. Steinkopff, Darmstadt, pp 139–148Google Scholar
  28. 28.
    Yandle T, Crozier I, Nicholls G, Espiner E, Carne A, Brennan S (1987) Amino acid sequence of atrial natriuretic peptides in human coronary sinus plasma. Biochem Biophys Res Commun 146:832–839Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • P. Schulz-Knappe
    • 1
  • K. Forssmann
    • 1
  • F. Herbst
    • 1
  • D. Hock
    • 1
  • R. Pipkorn
    • 1
  • W. G. Forssmann
    • 1
  1. 1.Institut für Anatomie und Zellbiologie IIIUniversität HeidelbergGermany

Personalised recommendations