Advertisement

Journal of Fluorescence

, Volume 24, Issue 6, pp 1803–1810 | Cite as

Interaction of Cu2+, Pb2+, Zn2+ with Trypsin: What is the Key Factor of their Toxicity?

  • Tong Zhang
  • Hao Zhang
  • Guiliang Liu
  • Canzhu Gao
  • Rutao Liu
ORIGINAL PAPER

Abstract

Heavy metals possess great endangerment to environment even human health because of their indissolubility and bioaccumulation. The toxicity of heavy metal ions (Cu2+, Pb2+, Zn2+) to trypsin was investigated by fluorescence, synchronous fluorescence, UV–vis absorption, circular dichroism (CD) spectroscopy, isothermal titration calorimetry (ITC), and enzyme activity assay. The experimental results showed that toxic effect of heavy metal ions was due to their own characteristic, rather than the electric charges of the ion. Zn2+ could not show the obvious toxicity to trypsin, while the structure and function of trypsin was damaged when the enzyme explored to Cu2+ and Pb2+. From the spectra results, we found that Cu2+ would bind with trypsin, which lead to the fluorescence quenched and hydrophobicity increased. Pb2+ could also change the structure and reduce the activity of trypsin in high concentration. In vitro measurement, the toxicity order of heavy metal ions to trypsin is: Cu2+ > Pb2+ > Zn2+. In addition, isothermal titration calorimetry analysis proved that the interactions between Cu2+, Pb2+, Zn2+ and trypsin were all spontaneous and exothermic, which indicated the adverse effect of these heavy metal ions to trypsin.

Keyword

Heavy metal ions Trypsin Spectroscopy Isothermal titration calorimetry 

Notes

Acknowledgments

This work is supported by NSFC (20875055, 21277081, 201477067), the Cultivation Fund of the Key Scientific and Technical Innovation Project, Research Fund for the Doctoral Program of Higher Education, Ministry of Education of China (708058, 20130131110016) are also acknowledged.

Supplementary material

10895_2014_1469_MOESM1_ESM.doc (2 mb)
ESM 1 (DOC 2065 kb)

References

  1. 1.
    Badugu R (2005) Fluorescence sensor design for transition metal ions: the role of the PIET interaction efficiency. J Fluoresc 15(1):71–83PubMedCrossRefGoogle Scholar
  2. 2.
    Tlili A et al (2011) Use of the MicroResp™ method to assess pollution-induced community tolerance to metals for lotic biofilms. Environ Pollut 159(1):18–24PubMedCrossRefGoogle Scholar
  3. 3.
    Berlizov AN et al (2007) Testing applicability of black poplar (Populus nigra L.) bark to heavy metal air pollution monitoring in urban and industrial regions. Sci Total Environ 372(2–3):693–706PubMedCrossRefGoogle Scholar
  4. 4.
    Jarup L (2003) Hazards of heavy metal contamination. Br Med Bull 68(1):167–182PubMedCrossRefGoogle Scholar
  5. 5.
    Chen TB (2005) Assessment of heavy metal pollution in surface soils of urban parks in Beijing, China. Chemosphere 60(4):542–551PubMedCrossRefGoogle Scholar
  6. 6.
    Agarwal SK (2009) Heavy metal pollution. APH Publ Vol 4Google Scholar
  7. 7.
    Wang Y et al (2007) The influence of soil heavy metals pollution on soil microbial biomass, enzyme activity, and community composition near a copper smelter. Ecotoxicol Environ Saf 67(1):75–81PubMedCrossRefGoogle Scholar
  8. 8.
    Muchuweti M et al (2006) Heavy metal content of vegetables irrigated with mixtures of wastewater and sewage sludge in Zimbabwe: implications for human health. Agric Ecosyst Environ 112(1):41–48CrossRefGoogle Scholar
  9. 9.
    Leung AO et al (2008) Heavy metals concentrations of surface dust from e-waste recycling and its human health implications in southeast China. Environ Sci Technol 42(7):2674–2680PubMedCrossRefGoogle Scholar
  10. 10.
    Yi Y, Yang Z, Zhang S (2011) Ecological risk assessment of heavy metals in sediment and human health risk assessment of heavy metals in fishes in the middle and lower reaches of the Yangtze River basin. Environ Pollut 159(10):2575–2585PubMedCrossRefGoogle Scholar
  11. 11.
    Meng, Z (2000) Environmental toxicology. China Environmental Science Press Vol. 8Google Scholar
  12. 12.
    Ghosh S (2008) Interaction of trypsin with sodium dodecyl sulfate in aqueous medium: a conformational view. Colloids Surf B: Biointerfaces 66(2):178–86PubMedCrossRefGoogle Scholar
  13. 13.
    Liu Y, Wang X (1995) Zinc, copper, copper, cadmium metal ions acute toxicity tests on the bayscallop spat. Aquat Sci 14:1–10Google Scholar
  14. 14.
    Fargasova A (2001) Phytotoxic effects of Cd, Zn, Pb, Cu and Fe on Sinapis alba L. seedlings and their accumulation in roots and shoots. Biol Plant 44(3):471–473CrossRefGoogle Scholar
  15. 15.
    Zhang H et al (2011) Toxic effects of different charged metal ions on the target—Bovine serum albumin. Spectrochim Acta A Mol Biomol Spectrosc 78(1):523–527PubMedCrossRefGoogle Scholar
  16. 16.
    Hu X, Yu Z, Liu R (2013) Spectroscopic investigations on the interactions between isopropanol and trypsin at molecular level. Spectrochim Acta A Mol Biomol Spectrosc 108:50–54PubMedCrossRefGoogle Scholar
  17. 17.
    Bosch Cabral C, Imasato H, Rosa JC (2002) Fluorescence properties of tryptophan residues in the monomeric d-chain of Glossoscolex paulistus hemoglobin: an interpretation based on a comparative molecular model. Biophys Chem 97(2):139–157PubMedCrossRefGoogle Scholar
  18. 18.
    Van De Weert M (2010) Fluorescence quenching to study protein-ligand binding: common errors. J Fluoresc 20(2):625–629PubMedCrossRefGoogle Scholar
  19. 19.
    Hemmila I, Laitala V (2005) Progress in lanthanides as luminescent probes. J Fluoresc 15(4):529–542PubMedCrossRefGoogle Scholar
  20. 20.
    F. Deng, Y. Wang, and J. Liu (2012) Fluorescence Spectroscopy interaction of copper ions with trypsin. 17th National Academic Conference on Molecular Spectroscopy.Google Scholar
  21. 21.
    Hong F et al (2003) Activity of the role of lead ions mechanism of trypsin. J Inorg Chem 19(2):129Google Scholar
  22. 22.
    Zhang H, Hao F, Liu R (2013) Interactions of lead (II) acetate with the enzyme lysozyme: a spectroscopic investigation. J Lumin 142:144–149CrossRefGoogle Scholar
  23. 23.
    Wang Y et al (2007) Experimental research on quantitative inversion models of suspended sediment concentration using remote sensing technology. Chin Geogr Sci 17(3):243–249CrossRefGoogle Scholar
  24. 24.
    HE JS et al (2011) Thermdynamics kinetics and structure chemistry. Acta Phys -Chim Sin 27(11):2499–2504Google Scholar
  25. 25.
    Geddes CD, Lakowicz JR (2002) Editorial: metal-enhanced fluorescence. J Fluoresc 12(2):121–129CrossRefGoogle Scholar
  26. 26.
    Ascenzi P et al (2003) The bovine basic pancreatic trypsin inhibitor (Kunitz inhibitor): a milestone protein. Curr Protein Pept Sci 4(3):231–251PubMedCrossRefGoogle Scholar
  27. 27.
    Prasher DC et al (1992) Primary structure of the Aequorea victoria green-fluorescent protein. Gene 111(2):229–233PubMedCrossRefGoogle Scholar
  28. 28.
    Amorín M, Castedo L, Granja JR (2003) New cyclic peptide assemblies with hydrophobic cavities: the structural and thermodynamic basis of a new class of peptide nanotubes. J Am Chem Soc 125(10):2844–2845PubMedCrossRefGoogle Scholar
  29. 29.
    Wang YQ, Chen TT, Zhang HM (2010) Investigation of the interactions of lysozyme and trypsin with biphenol a using spectroscopic methods. Spectrochim Acta A Mol Biomol Spectrosc 75(3):1130–1137PubMedCrossRefGoogle Scholar
  30. 30.
    Tsao MS et al (2005) Erlotinib in lung cancer—molecular and clinical predictors of outcome. N Engl J Med 353(2):133–144PubMedCrossRefGoogle Scholar
  31. 31.
    Sui Y et al (2007) Catalytic and asymmetric Friedel–crafts alkylation of indoles with nitroacrylates. Appl Synth Tryptophan Analogues Tetrahedron 63(24):5173–5183Google Scholar
  32. 32.
    Kelly SM, Jess TJ, Price NC (2005) How to study proteins by circular dichroism. Biochim Biophys Acta (BBA)-Proteins Proteomics 1751(2):119–139CrossRefGoogle Scholar
  33. 33.
    Chai J et al (2013) Investigation on potential enzyme toxicity of clenbuterol to trypsin. Spectrochim Acta A Mol Biomol Spectrosc 105:200–206PubMedCrossRefGoogle Scholar
  34. 34.
    Dullweber F et al (2001) Factorising ligand affinity: a combined thermodynamic and crystallographic study of trypsin and thrombin inhibition. J Mol Biol 313(3):593–614PubMedCrossRefGoogle Scholar
  35. 35.
    Turnbull WB, Daranas AH (2003) On the value of c: can low affinity systems be studied by isothermal titration calorimetry? J Am Chem Soc 125(48):14859–14866PubMedCrossRefGoogle Scholar
  36. 36.
    Vermeirssen V, Van Camp J, Verstraete W (2002) Optimisation and validation of an angiotensin-converting enzyme inhibition assay for the screening of bioactive peptides. J Biochem Biophys Methods 51(1):75–87PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.School of Environmental Science and Engineering, China -America CRC for Environment & Health, Shandong ProvinceShandong UniversityJinanPeople’s Republic of China
  2. 2.Shandong Institute for Food and Drug ControlJinanChina

Personalised recommendations