Skip to main content
SpringerLink
Log in

Study on dipeptidylpeptidase II (DPP II)

  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

The activity of dipeptidylpeptidase II (DPP II; E.C. 3.4.14.2) was investigated by biochemical and histochemical methods in rat, mouse and guinea-pig organs as well as in human enterobiopsies. Lys-Pro-MNA and Ala-Pro-MNA showed the most favorable kinetic properties (K m , V max) and proved to be the most sensitive substrates for biochemical and histochemical studies of DPP II. Lys-Ala-MNA is more specific and is to be preferred due to its relatively low hydrolysis by DPP IV. Lys-Ala-2NA is suitable for the biochemical determination of DPP II activity. Lys-Ala-1NA, Leu-Ala-2NA, Phe-Pro-2NA and Phe-Pro-MNA are inferior. The pH optimum of DPP II amounts to 5.5. Cacodylate, phosphate, citric acid phosphate and succinate buffers deliver similar hydrolysis rates; with citrate and acetate buffers the recorded activities are lower. The reaction can be inhibited by 1 mM DFP, 50 mM Tris and 10 mM puromycin. In the ileum of suckling rats and in human enterobiopsies similar data (K m , pH optimum, optimal substrate concentration) were obtained by biochemical determination and by quantitative histochemistry (microdensitometry) with Lys-Ala-MNA. For the histochemical demonstration of DPP II freeze-dried celloidin-coated cryostat sections are very suitable. Frozen sections of formaldehyde and glutaraldehyde fixed tissue blocks are inferior due to a higher inhibition of DPP II and less precise localization of the azo-dye. K m values and optimal pH are identical in fresh and fixed material. Fast Blue B is the best coupling agent for light microscopical localization. DPP II is present in all organs and tissues investigated. Conspicious organ and species differences exist. In adult rats the highest DPP II activity resides in the kidney, epididymis and spleen; in guinea-pigs the epididymis and testis are the most active organs. In the majority of guinea-pig organs the DPP II activity is lower than in rats. The histochemical demonstration of DPP II shows, in addition, cell-dependent differences of DPP II activity. In most cells the enzyme activity is depicted in lysosomes. Highly active are lysosomes of cells of proximal renal tubules, macrophages, thyroid cells, clear and principal cells of the epididymis of adult animals and of enterocytes of suckling rats. Lysosomes of endocrine cells of adenohypophysis, pancreaas, stomach, small intestine and nerve cells display moderate activity. In lysosomes of smooth muscle cells (intestine, myometrium), myocardial cells, and fibers of striated muscle the enzyme is also present. Spermatids and sperms of guinea-pigs are highly active. In some cases secretion granules of endocrine and exocrine gland cells display a positive reaction. Possibly the Golgi apparatus and the endoplasmic reticulum also show a positive staining in the principle cells of the rat and mouse epididymis. Furthermore, DPP II seems to be secreted into the lumen of several organs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Afting EG (1979) Activity of a rat uterus proteinase inhibitor during pregnancy and involution. Its possible importance in control of proteolysis in the myometrium. In: Holzer H, Tschesche H (eds) Biological functions of proteinases. Springer, Berlin Heidelberg New York, pp 87–93

    Google Scholar 

  • Blenk R, Gossrau R (1980) Biochemische und quantitativ-histochemische Untersuchungen von Proteinasen in der Gld. submandibularis von Ratten. Acta Histochem Suppl 24 (in press)

  • Dayton WR, Goll DE, Stromer MH, Reville WJ, Zeece MG, Robson RM (1975) Some properties of a Ca2+-activated protease that may be involved in myofibrillar protein turnover. In: Reich E, Rifkin DB, Shaw E (eds) Proteases and biological control. Cold Spring Harbor, New York, pp 551–577

  • Gossrau R (1977a) Ageing and death of intestinal epithelial cells during pre- and postnatal development. Proc R Microsc Soc 12:216

    Google Scholar 

  • Gossrau R (1977b) Peptidasen I. Histochemische Untersuchungen mit 2-Naphthylamiden und Hexazonium-p-rosanilin. Histochemistry 54:311–330

    Google Scholar 

  • Gossrau R (1978) Histochemische und biochemische Untersuchungen von Peptidasen mit Aminosäure-und Peptid-2-naphthylamiden. Acta Histochem (Suppl) 20:279–285

    Google Scholar 

  • Gossrau R (1979a) Peptidasen II. Zur Lokalisation der Dipeptidylpeptidase IV (DPP IV). Histochemische und biochemische Untersuchung. Histochemistry 60:231–248

    Google Scholar 

  • Gossrau R (1979b) Biochemistry and histochemistry of lysosomal exo- and endopeptidases. Commun Czech Soc Histochem Cytochem 7:22

    Google Scholar 

  • Gossrau R (1980a) Zur Eignung verschiedener Diazoniumsalze für histochemische Peptidasenuntersuchungen. Acta Histochem (Suppl) 21:229

    Google Scholar 

  • Gossrau R (1980b) Conventional techniques for membrane-bound enzymes. In: Trends in enzyme histochemistry and cytochemistry (Ciba Foundation Symposion 73). Excerpta Medica, Amsterdam, pp 67–80

    Google Scholar 

  • Gossrau R (1980c) Zur Lokalisation lysosomaler Peptidasen in Labornagerorganen. Anat Anz 147:497–498

    Google Scholar 

  • Gossrau R (1980d) Peptidasen: Neue Einblicke in Organfunktionen. Verh Anat Ges 74 (in press)

  • Gossrau R (1980e) Investigation of proteinases in the digestive tract using 4-methoxy-2-naphthylamine (MNA) substrates. J Histochem Cytochem (in press)

  • Gossrau R (1980f) Proteinases in the secretory, resorptive and immune system of the digestive tract. Histochemical Society, 31st Annual Meeting, April, 11–15, New Orleans

  • Hamilton DW (1975) Structure and function of the epithelium lining the ductuli efferentes, ductus epididymidis and ductus deferens in the rat. In: Hamilton DW, Greep RO (eds) Handbook of physiology, vol 5. Endocrinology, Section 7. Male reproductive system. Am Physiol Soc, Bethesda, pp 259–301

    Google Scholar 

  • Hartmann K, Gossrau R (1978) Die postnatale Entwicklung des Dünndarmepithels beim Meerschweinchen. Prog Histochem Cytochem 11: Heft 2, 1–67

    Google Scholar 

  • Kenny AJ (1977) Proteinases associated with cell membranes. In: Barrett AJ (ed) Proteinases in mammalian cells and tissues. North-Holland, Amsterdam New York Oxford, pp 393–444

    Google Scholar 

  • Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am chem Soc 56:658–666

    Google Scholar 

  • Lojda Z (1975) The use of hexazonium-p-rosaniline in the histochemical demonstration of peptidases. Histochemistry 44:323–335

    Google Scholar 

  • Lojda Z (1977) Studies on dipeptidyl(amino)peptidase IV (glycylproline naphthylamidase). I. Lymphocytes. Histochemistry 54:299–309

    Google Scholar 

  • Lojda Z (1979a) Studies on dipeptidyl(amino)peptidase IV (glycylproline naphthylamidase). II. Blood vessels. Histochemistry 59:153–166

    Google Scholar 

  • Lojda Z (1979b) The histochemical demonstration of brush border endopeptidase. Histochemistry 64:205–221

    Google Scholar 

  • Lojda Z, Gossrau R (1980a) Study on aminopeptidase A. Histochemistry 67:267–290

    Google Scholar 

  • Lojda Z, Gossrau R (1980b) Distribution of peptidases in the central nervous system (manuscript in preparation)

  • Lojda Z, Gossrau R, Schiebler TH (1976) Enzymhistochemische Methoden. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Lojda Z, Gossrau R, Schiebler TH (1979a) Enzyme histochemistry. A laboratory manual. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Lojda Z, Heřmansky F, Benešová E, Šálková J, Lodrová V (1979b) Dipeptidylaminopeptidase IV in lymphocytes of peripheral blood of man and the importance of its evidence in malignant lymphomas. Sb Lek 81:200–207

    Google Scholar 

  • McDonald JK, Callahan PX, Ellis S, Smith RE (1971) Polypeptide degradation by dipeptidylaminopeptidase I (cathepsin C) and related peptidases. In: Barrett AJ, Dingle TJ (eds) Tissue proteinases. North-Holland, Amsterdam New York Oxford, pp 69–107

    Google Scholar 

  • McDonald JK, Leibach RE, Grindeland RE, Ellis S (1968a) Purification of dipeptidylaminopeptidase II (dipeptidylarylamidase II) of the anterior pituitary gland; peptidase and dipeptidase esterase activities. J Biol Chem 243:4143–4150

    Google Scholar 

  • McDonald JK, Reilly TJ, Zeitman BB, Ellis S (1968b) Dipeptidylarylamidase II of the pituitary; properties of lysyslalanyl-β-naphthylamide hydrolysis, inhibition by cations, distribution in tissues and subcellular localization. J Biol Chem 243:2028–2037

    Google Scholar 

  • McDonald JK, Schwabe C (1977) Intracellular exopeptidases. In: Barrett AJ (ed) Proteinases in mammalian cells and tissues. North-Holland, Amsterdam New York Oxford, pp 311–391

    Google Scholar 

  • Morton D (1977) The occurrence and function of proteolytic enzymes in the reproductive tract of mammals. In: Barrett AJ (ed) Proteinases in mammalian cells and tissues. North-Holland, Amsterdam New York Oxford, pp 445–500

    Google Scholar 

  • Nachlas MM, Goldstein TP, Rosenblatt DH, Kirsch M, Seligman AM (1959) Influence of chemical structure on the rate of azocoupling and its significance in histochemical methodology. J Histochem Cytochem 7:50–65

    Google Scholar 

  • Neurath H (1975) Limited proteolysis and zymogen activation. In: Reich E, Rifkin DB, Shaw E (eds) Proteases and biological control. Cold Spring Harbor, New York, pp 51–64

  • Pennington RJT (1977) Proteinases of muscle. In: Barrett AJ (ed) Proteinases in mammalian cells and tissues. North-Holland, Amsterdam New York Oxford, pp 515–543

    Google Scholar 

  • Sannes PL, McDonald JK, Allen RC, Spicer SS (1979) Cytochemical localization and biochemical characterization of dipeptidylaminopeptidase II in macrophages and mast cells. J Histochem Cytochem 27:1496–1498

    Google Scholar 

  • Sannes PL, Schoefield BH, Coleman RA, McDonald DF (1980) Localization of dipeptidylaminopeptidase in connective tissue cells and macrophages. Histochemical Society, 31st Annual Meeting, April 11–15, New Orleans

  • Schröder J, Gossrau R (1980) Ultracytochemical demonstration of proteinases with hexazotized new fuchsine. Acta Histochem Suppl 25 (in press)

  • Smith RE, Dean PN (1979) A study of acid phosphatase and dipeptidylaminopeptidase II in monodispersed anterior pituitary cells using flow cytometry and electron microscopy. J Histochem Cytochem 27:1499–1504

    Google Scholar 

  • Smith RE, van Frank RM (1975) The use of amino acid derivatives of 4-methoxy-2-naphthylamine for the assay and subcellular localization of tissue proteinases. In: Dingle JT, Dean RJ (eds) Lysosomes in biology and pathology, vol IV. North-Holland, Amsterdam New York Oxford, pp 193–249

    Google Scholar 

  • Steiner DF, Kemmler W, Tager HS, Rubenstein AH, Lernmark A, Zühlke H (1975) Proteolytic mechanisms in the biosynthesis of polypeptide hormones. In: Reich E, Rifkin DB, Shaw E (eds) Proteases and biological control. Cold Spring Harbor, New York, pp 531–549

  • Williams-Ashman HG (1975) Introductory overview of the participation of proteinases and their regulators in mammalian reproductive physiology. In: Reich E, Rifkin DB, Shaw E (eds) Proteinases and biological control. Cold Spring Harbor, New York, pp 677–681

  • Zunke U, Gossrau R (1980a) Zur Morphologie und Zonengliederung des Nebenhodenganges der Ratte. I. Semidünnschnittuntersuchungen. Anat Anz (in press)

  • Zunke U, Gossrau R (1980b) Zur Morphologie und Zonengliederung des Nebenhodenganges der Ratte. II. Elektronenmikroskopische Untersuchungen. Anat Anz (in press)

Download references

Author information

Authors and Affiliations

  1. Department of Anatomy, University of Würzburg, Koellikerstrasse 6, D-8700, Würzburg, Federal Republic of Germany

    R. Gossrau

  2. Laboratory of Histochemistry, Faculty of Medicine, Charles University, Studničkova 2, ČS-12800, Prague 2, ČSSR

    Z. Lojda

Authors
  1. R. Gossrau
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Z. Lojda
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supported by Deutsche Forschungsgemeinschaft (SFB 105)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gossrau, R., Lojda, Z. Study on dipeptidylpeptidase II (DPP II). Histochemistry 70, 53–76 (1980). https://doi.org/10.1007/BF00508846

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00508846

Keywords

Search

Navigation