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Histochemical Demonstration of Tripeptidyl Aminopeptidase I

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Histochemistry of Single Molecules

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1560))

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Abstract

Enzyme histochemical methods are valuable for the studies on the enzyme involvement in different pathological processes. Here we describe two protocols for chromogenic and fluorogenic histochemical demonstration of tripeptidyl aminopeptidase I (TPPI), a protease that is crucial for neuronal functions. The procedures are based on newly synthesized substrates for TPPI—glycyl-l-prolyl-l-metionyl-5-chloro-1-anthraquinonylhydrazide (GPM-CAH) and glycyl-l-prolyl-l-metionyl-4-hydrazido-N-hexyl-1,8-naphthalimide (GPM-HHNI). Using such protocols, precise enzyme localization can be obtained in tissue sections of mammalian organs.

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References

  1. Wlodawer A, Li M, Gustchina A, Oyama H et al (2003) Structural and enzymatic properties of the sedolisin family of serine-carboxyl peptidases. Acta Biochim Polon 50:81–102

    CAS  PubMed  Google Scholar 

  2. Ezaki J, Takeda-Ezaki M, Oda K et al (2000) Characterization of endopeptidase activity of tripeptidyl peptidase-I/CLN2 protein which is deficient in classical late infantile neuronal ceroid lipofuscinosis. J Biochem Biophys Res Commun 268:904–908

    Article  CAS  Google Scholar 

  3. Wlodawer A, Durell SR, Li M et al (2003) A model of tripeptidyl peptidase I (CLN2), a ubiquitous and highly conserved member of the sedolisin family of serine-carboxyl peptidases. BMC Struct Biol 3:8–18

    Article  PubMed  PubMed Central  Google Scholar 

  4. Sharp JD, Wheeler RB, Lake BD et al (1997) Loci for classical and a variant late infantile neuronal ceroid lipofuscinosis map to chromosomes 11p15 and 15q21-23. Hum Mol Genet 6:591–595

    Article  CAS  PubMed  Google Scholar 

  5. Lin L, Lobel P (2001) Production and characterization of recombinant human CLN2 protein for enzyme-replacement therapy in late infantile neuronal ceroid lipofuscinosis. Biochem J 357:49–55

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Page AE, Fuller K, Chambers TJ et al (1993) Purification and characterization of a tripeptidyl peptidase I from human osteoclastomas: evidence for its role in bone resorption. Arch Biochem Biophys 306:354–359

    Article  CAS  PubMed  Google Scholar 

  7. Warburton MJ, Bernardini F (2001) The specificity of lysosomal tripeptidyl peptidase-I determined by its action on angiotensin-II analogues. FEBS Lett 500:145–148

    Article  CAS  PubMed  Google Scholar 

  8. Kopan S, Sivasubramaniam U, Warburton MJ (2004) The lysosomal degradation of neuromedin B is dependent on tripeptidyl peptidase-I: evidence for the impairment of neuropeptide degradation in late-infantile neuronal ceroid lipofuscinosis. Biochem Biophys Res Commun 319:58–65

    Article  CAS  PubMed  Google Scholar 

  9. Junaid M, Wu G, Pullarkat R (2000) Purification and characterization of bovine brain lysosomal pepstatin-insensitive proteinase, the gene product deficient in the human late-infantile neuronal ceroid lipofuscinosis. J Neurochem 74:287–294

    Article  CAS  PubMed  Google Scholar 

  10. Bernardini F, Warburton MJ (2002) Lysosomal degradation of cholecystokinin-(29–33)-amide in mouse brain is dependent on tripeptidyl peptidase-I: implications for the degradation and storage of peptides in classical late-infantile neuronal ceroid lipofuscinosis. Biochem J 366:521–529

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Sleat DE, Donnelly JR, Lackland H et al (1997) Association of mutations in a lysosomal protein with classical late-infantile neuronal ceroid lipofuscinosis. Science 277:1802–1805

    Article  CAS  PubMed  Google Scholar 

  12. Golabek A, Kida E (2006) Tripeptidyl-peptidase I in health and disease. Biol Chem 387:1091–1099

    Article  CAS  PubMed  Google Scholar 

  13. Junaid MA, Clark GM, Pullarkat RK (2000) A lysosomal pepstatin-insensitive proteinase as a novel biomarker for breast carcinoma. Int J Biol Markers 15:129–134

    CAS  PubMed  Google Scholar 

  14. Altorjay A, Paal B, Sohar N et al (2005) Significance and prognostic value of lysosomal enzyme activities measured in surgically operated adenocarcinomas of the gastroesophageal junction and squamous cell carcinomas of the lower third of esophagus. World J Gastroenterol 11:5751–5756

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Tsukamato T, Lida J, Dobashi Y (2006) Overexpression in colorectal carcinoma of two lysosomal enzymes, CLN2 and CLN1, involved in neuronal ceroid lipofuscinosis. Cancer 106:1489–1497

    Article  Google Scholar 

  16. Autefage H, Albinet V, Garcia V et al (2009) Lysosomal serine protease CLN2 regulates tumor necrosis factor-alfa-mediated apoptosis in a Bid-dependent manner. J Biol Chem 284:11507–11516

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Van Beersel G, Tihon E, Demine S et al (2013) Different molecular mechanisms involved in spontaneous and oxidative stress-induced mitochondrial fragmentation in tripeptidyl peptidase-1 (TPP-1)-deficient fibroblasts. Biosci Rep 33:243–258

    Google Scholar 

  18. Koike M, Shibata M, Ohsawa Y et al (2002) The expression of trypeptidyl peptidase I in various tissues of rats and mice. Arch Histol Cytol 65:219–232

    Article  CAS  PubMed  Google Scholar 

  19. Yayoi Y, Ohsawa Y, Koike M et al (2001) Specific localization of lysosomal aminopeptidases in type II alveolar epithelial cells of the rat lung. Arch Histol Cytol 64:89–97

    Article  CAS  PubMed  Google Scholar 

  20. Kurachi Y, Oka A, Itoh M et al (2001) Distribution and development of CLN2 protein, the late-infantile neuronal ceroid lipofuscinosis gene product. Acta Neuropathol 102:20–26

    CAS  PubMed  Google Scholar 

  21. Kida E, Golabeki A, Walus M et al (2001) Distribution of trypeptidyl peptidase I in human tissues under normal and pathological conditions. J Neuropathol Exp Neurol 60:280–292

    Article  CAS  PubMed  Google Scholar 

  22. Dikov A, Dimitrova M, Ivanov I et al (2000) Original method for the histochemical demonstration of tripeptidyl aminopeptidase I. Cell Mol Biol 46:1219–1225

    CAS  PubMed  Google Scholar 

  23. Ivanov I, Tascheva D, Todorova R et al (2009) Synthesis and use of 4-peptidylhydrazido-N-hexyl-1,8-naphthalimides as fluorogenic histochemical substrates for dipeptidyl peptidase IV and tripeptidyl peptidase I. Eur J Med Chem 44:384–392

    Article  CAS  PubMed  Google Scholar 

  24. Dimitrova M, Ivanov I, Deleva D (2009) Distribution of tripeptidyl peptidase I activity in the rat brain and spinal cord. Compt Rend Acad Bulg Sci 62:729–734

    CAS  Google Scholar 

  25. Dimitrova M, Ivanov I, Todorova R et al (2008) Fluorescent localization of tripeptidyl peptidase I activity in tissue sections of Balb/c mice reproductive organs. Compt Rend Acad Bulg Sci 61:349–356

    CAS  Google Scholar 

  26. Atanasova D, Lazarov N (2015) Histochemical demonstration of tripeptidyl aminopeptidase I in the rat carotid body. Acta Histochem 117:219–222

    Article  CAS  PubMed  Google Scholar 

  27. Dimitrova M, Deleva D (2009) Histochemical study of the changes in trypeptidyl peptidase I activity in developing rat brain and spinal cord. Compt Rend Acad Bulg Sci 62:1407–1412

    CAS  Google Scholar 

  28. Suopanki J, Partanen S, Ezaki J et al (2000) Developmental changes in the expression of neuronal ceroid lipofuscinoses-linked proteins. Mol Genet Metab 71:190–194

    Article  CAS  PubMed  Google Scholar 

  29. Dimitrova M, Deleva D, Pavlova V et al (2011) Developmental study of tripeptidyl peptidase I activity in the mouse central nervous system and peripheral organs. Cell Tissue Res 346:141–149

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The authors greatly thank Prof Dr. Michail Davidoff for protocol consultation.

Author information

Authors and Affiliations

  1. Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Sofia, Bulgaria

    Mashenka B. Dimitrova

  2. Institute of Neurobiology, Bulgarian Academy of Sciences, Sofia, Bulgaria

    Dimitrinka Y. Atanasova & Nikolai E. Lazarov

  3. Department of Anatomy, Faculty of Medicine, Trakia University, Stara Zagora, Bulgaria

    Dimitrinka Y. Atanasova

  4. Department of Anatomy and Histology, Medical University of Sofia, 2, Zdrave Street, 1431, Sofia, Bulgaria

    Nikolai E. Lazarov

Authors
  1. Mashenka B. Dimitrova
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  2. Dimitrinka Y. Atanasova
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  3. Nikolai E. Lazarov
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Corresponding author

Correspondence to Nikolai E. Lazarov .

Editor information

Editors and Affiliations

  1. Department of Biology and Biotechnology, University of Pavia, Pavia, Italy

    Carlo Pellicciari

  2. Department of Biology and Biotechnology, University of Pavia, Pavia, Italy

    Marco Biggiogera

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Dimitrova, M.B., Atanasova, D.Y., Lazarov, N.E. (2017). Histochemical Demonstration of Tripeptidyl Aminopeptidase I. In: Pellicciari, C., Biggiogera, M. (eds) Histochemistry of Single Molecules. Methods in Molecular Biology, vol 1560. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6788-9_4

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  • DOI: https://doi.org/10.1007/978-1-4939-6788-9_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6787-2

  • Online ISBN: 978-1-4939-6788-9

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