Skip to main content
SpringerLink
Log in

Kinetics of Alloxan-Induced Inhibition on δ-Aminolevulinate Dehydratase Activity in Mouse Liver Homogenates

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

This study evaluated the effects of alloxan on the kinetics properties of the δ-aminolevulinate dehydratase (δ-ALA-D) using mouse liver homogenates. δ-ALA-D is an important sulfhydryl enzyme that catalyses the second step in heme biosynthesis and is commonly diminished in experimental and human diabetes. Despite the known effects of alloxan in models of experimental diabetes, there are no data in the literature demonstrating the effects of alloxan on the kinetics properties of the δ-ALA-D. The results showed that alloxan (1.25–20 μM) caused a concentration-dependent inhibition of hepatic δ-ALA-D activity. The inhibition constant (K i ) for alloxan-induced inhibition on δ-ALA-D was 3.64 μM. The alloxan (5 μM) caused a decrease in V max (65.8%) and in K m (53.1%), which is suggestive of an uncompetitive inhibition of enzyme. In addition, dithiothreitol (700 and 1,000 μM) completely prevented the δ-ALA-D activity inhibition induced by 10 and 20 μM alloxan. Similar protection was obtained in the presence of 2,000 μM glutathione. Therefore, this work showed that the inhibition of hepatic δ-ALA-D activity can be obtained in vitro at low micromolar levels of alloxan, and can also be prevented by reducing agents. Moreover, these results may help to understand the abnormalities in heme pathway found in models of experimental diabetes in vivo.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Sassa, S. (1998). Seminars in Liver Disease, 18, 5–101.

    Article  Google Scholar 

  2. Sterling, K., Silver, M., & Ricketts, H. T. (1949). Archives of Internal Medicine, 84, 965–975.

    Article  CAS  Google Scholar 

  3. Burnham, T. K., & Fosnaugh, R. P. (1961). Archives of Dermatology, 83, 717–722.

    Article  CAS  Google Scholar 

  4. Stein, J. A., & Tschudy, D. P. (1970). Medicine (Baltimore), 49, 1–16.

    CAS  Google Scholar 

  5. Goerz, G., & Korda, S. (1977). Zeits Hautkrank, 52, 1165–1174.

    CAS  Google Scholar 

  6. Székely, E., Bor, M., Tasnádi, G., Várnai, K., Almási, A., & Blázovic, A. (2006). Clinical Hemorheology and Microcirculation, 35, 387–396.

    Google Scholar 

  7. Grossman, M. E., Bickers, D. R., Poh-Fitzpatrick, M. B., Deleo, V. A., & Harber, L. C. (1979). American Journal of Medicine, 67, 277–286.

    Article  CAS  Google Scholar 

  8. Lisi, P., Santeusanio, F., & Lombardi, G. (1983). Dermatologica, 166, 287–293.

    Article  CAS  Google Scholar 

  9. Sibbald, R. G., & Schachter, R. K. (1984). International Journal of Dermatology, 23, 567–584.

    Article  CAS  Google Scholar 

  10. Mengistu, M. (1987). Tropical and Geographical Medicine, 39, 361–365.

    CAS  Google Scholar 

  11. Magnin, P. H., & Lenczner, J. M. (1990). Argentina de Dermatolologia, 71, 45–47.

    Google Scholar 

  12. Santamaría, V. G., & Barrios, E. G. (2001). Revista del Centro Dermatológico Pascua, 10, 1–12.

    Google Scholar 

  13. Caballero, F. A., Gerez, E. N., Polo, C. F., Vazquez, E. S., & Batlle, A. M. C. (1998). General Pharmacology, 31, 441–445.

    Article  CAS  Google Scholar 

  14. Folmer, V., Soares, J. C. M., & Rocha, J. B. T. (2002). International Journal of Biochemistry and Cell Biology, 34, 1279–1285.

    Article  CAS  Google Scholar 

  15. Folmer, V., Soares, J. C. M., Gabriel, D., & Rocha, J. B. T. (2003). Journal of Nutrition, 133, 2165–2170.

    CAS  Google Scholar 

  16. Brito, V. B., Folmer, V., Soares, J. C., Silveira, I. D., & Rocha, J. B. (2007). Nutrition, 23, 818–826.

    Article  CAS  Google Scholar 

  17. Brito, V. B., da Rocha, J. B. T., Puntel, G. O., da Luz, S. C. A., Barbosa, N. B. V., de Carvalho, N. R., et al. (2011). Experimental and Toxicologic Pathology, 63, 443–451.

    Article  CAS  Google Scholar 

  18. Sassa, S. (1982). Enzyme, 28, 133–145.

    CAS  Google Scholar 

  19. Shemim, D. (1976). Philosophical Transactions of the Royal Society of London, Series B, 273, 109–115.

    Article  Google Scholar 

  20. Tsukamoto, I., Yoshinaga, T., & Sano, S. (1979). Biochimica et Biophysica Acta, 570, 167–178.

    CAS  Google Scholar 

  21. Farina, M., Brandão, R., Lara, F. S., Soares, F. A., Souza, D. O., & Rocha, J. B. T. (2003). Toxicology Letters, 139, 55–66.

    Article  CAS  Google Scholar 

  22. Perottoni, J., Meotti, F. C., Folmer, V., Pivetta, L., Nogueira, C. W., Zeni, G., et al. (2005). Environmental Toxicology and Pharmacology, 19, 239–248.

    Article  CAS  Google Scholar 

  23. Spencer, P., & Jordan, P. M. (1994). Biochemical Journal, 300, 373–381.

    CAS  Google Scholar 

  24. Spencer, P., & Jordan, P. M. (1995). Biochemical Journal, 305, 151–158.

    CAS  Google Scholar 

  25. Valentini, J., Grotto, D., Paniz, C., Roehrs, M., Burg, G., & Garcia, S. C. (2008). Biomedicine & Pharmacotherapy, 62, 378–382.

    Article  CAS  Google Scholar 

  26. Dunn, J. S., Sheehan, H. L., & McLetchie, N. G. B. (1943). Lancet, 1, 484–487.

    Google Scholar 

  27. Lenzen, S., & Panten, U. (1988). Diabetologia, 31, 337–342.

    Article  CAS  Google Scholar 

  28. Grankvist, K., Marklund, S. L., Sehlin, J., & Taljedal, I. B. (1979). Biochemical Journal, 182, 17–25.

    CAS  Google Scholar 

  29. Nukatsuka, M., Sakurai, H., & Kawada, J. (1989). Biochemical and Biophysical Research Communications, 165, 278–283.

    Article  CAS  Google Scholar 

  30. Elsner, M., Gurgul-Convey, E., & Lenzen, S. (2008). Antioxidants and Redox Signaling, 10, 691–699.

    Article  CAS  Google Scholar 

  31. Lenzen, S. (2008). Diabetologia, 51, 216–226.

    Article  CAS  Google Scholar 

  32. Munday, R. (1988). Biochemical Pharmacology, 37, 409–413.

    Article  CAS  Google Scholar 

  33. Elsner, M., Gurgul-Convey, E., & Lenzen, S. (2006). Free Radical Biology and Medicine, 41, 825–834.

    Article  CAS  Google Scholar 

  34. Winterbourn, C. C., & Munday, R. (1989). Biochemical Pharmacology, 38, 271–277.

    Article  CAS  Google Scholar 

  35. Ammon, H. P., & Mark, M. (1985). Cell Biochemistry Function, 3, 157–171.

    Article  CAS  Google Scholar 

  36. Boquist, L., & Ericsson, I. (1984). FEBS Letters, 178, 245–248.

    Article  CAS  Google Scholar 

  37. Tiedge, M., Krug, U., & Lenzen, S. (1997). Biochimica et Biophysica Acta, 1337, 175–190.

    Article  CAS  Google Scholar 

  38. Zhang, X., Liang, W., Mao, Y., Li, H., Yang, Y., & Tan, H. (2009). Biomedicine & Pharmacotherapy, 63, 180–186.

    Article  CAS  Google Scholar 

  39. Barbosa, N. B., Rocha, J. B. T., Zeni, G., Emanuelli, T., Beque, M. C., & Braga, A. L. (1998). Toxicology and Applied Pharmacology, 149, 243–253.

    Article  CAS  Google Scholar 

  40. Bradford, M. M. (1976). Analytical Biochemistry, 72, 248–254.

    Article  CAS  Google Scholar 

  41. Elsner, M., Tiedge, M., Guldbakke, B., Munday, R., & Lenzen, S. (2002). Diabetologia, 45, 1542–1549.

    Article  CAS  Google Scholar 

  42. Leech, R., & Bailey, C. (1945). Journal of Biological Chemistry, 157, 525–542.

    CAS  Google Scholar 

  43. Bruckmann, G., & Wertheimer, E. (1947). Journal of Biological Chemistry, 168, 241–256.

    CAS  Google Scholar 

  44. Bhattacharya, S. K., Robson, J. S., & Stewart, C. P. (1956). Biochemical Journal, 62, 12–21.

    CAS  Google Scholar 

  45. Kiersztan, A., Baranska, A., Hapka, M., Lebiedzinska, M., Winiarska, K., Dudziak, M., et al. (2009). Chemico-Biological Interactions, 177, 161–171.

    Article  CAS  Google Scholar 

  46. Szkudelski, T., Kandulska, K., & Okulicz, M. (1998). Physiological Research, 47, 343–346.

    CAS  Google Scholar 

  47. Grankvist, K., & Marklund, S. L. (1986). International Journal of Biochemistry, 18, 109–113.

    Article  CAS  Google Scholar 

  48. Zhang, H., Gao, G., & Brunk, U. T. (1992). Acta Pathologica Microbiologica et Immunologica Scandinavica, 100, 317–325.

    Article  CAS  Google Scholar 

  49. Patterson, J. W., Lazarow, A., & Levey, S. (1949). Journal of Biological Chemistry, 177, 197–204.

    CAS  Google Scholar 

  50. Caballero, F., Gerez, E., Pólo, C., Mompo, O., Vázquez, E., Schultz, R., et al. (1995). Medicina (Buenos Aires), 55, 117–124.

    CAS  Google Scholar 

  51. Czyzyk, A., & Gregor, A. (1971). Diabetologia, 152, 152–155.

    Article  Google Scholar 

  52. Bitar, M., & Weiner, M. (1984). Diabetes, 33, 37–44.

    Article  CAS  Google Scholar 

  53. Djordjevic, V. (1985). Archives Internationales de Physiologie et de Biochimie, 93, 285–290.

    Article  CAS  Google Scholar 

  54. Mohamed, A. K., Bierhaus, A., Schiekofer, S., Tritschler, H., Ziegler, R., & Nawroth, P. P. (1999). Biofactors, 10, 157–167.

    Article  CAS  Google Scholar 

  55. Wolff, S. P., & Dean, R. T. (1987). Biochemical Journal, 245, 243–250.

    CAS  Google Scholar 

  56. Fernández-Cuartero, B., Rebollar, J. L., Batlle, A., & Salamanca, R. E. (1999). International Journal of Biochemistry and Cell Biology, 31, 479–488.

    Article  Google Scholar 

  57. Tippett, P. S., & Neet, K. E. (1983). Archives of Biochemistry and Biophysics, 222, 285–298.

    Article  CAS  Google Scholar 

  58. Lazarow, A. (1949). Physiological Reviews, 29, 48–74.

    CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by FAPERGS (Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul), CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior), and CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico).

Author information

Authors and Affiliations

  1. Departamento de Química, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, UFSM, 97105-900, Santa Maria, Rio Grande do Sul, Brazil

    Verônica B. Brito, Vanderlei Folmer & João Batista T. Rocha

  2. Centro de Química e Bioquímica, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, UL, C8/8.4.61, Campo Grande, 1749-016, Lisbon, Portugal

    Verônica B. Brito

  3. Universidade Federal do Pampa, UNIPAMPA, Campus de Uruguaiana, BR 472 - KM 592, CEP 97500-970, Uruguaiana, Rio Grande do Sul, Brazil

    Vanderlei Folmer

  4. Faculdade de Farmácia, Universidade de Lisboa, UL, Avenida Doutor Gama Pinto, 1649-003, Lisbon, Portugal

    Inês Isabel R. Guerra

Authors
  1. Verônica B. Brito
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vanderlei Folmer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Inês Isabel R. Guerra
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. João Batista T. Rocha
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Verônica B. Brito.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Brito, V.B., Folmer, V., Guerra, I.I.R. et al. Kinetics of Alloxan-Induced Inhibition on δ-Aminolevulinate Dehydratase Activity in Mouse Liver Homogenates. Appl Biochem Biotechnol 166, 1047–1056 (2012). https://doi.org/10.1007/s12010-011-9492-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-011-9492-8

Keywords

Search

Navigation