Advertisement

Archives of Toxicology

, Volume 86, Issue 11, pp 1693–1702 | Cite as

S-Mercuration of rat sorbitol dehydrogenase by methylmercury causes its aggregation and the release of the zinc ion from the active site

  • Hironori Kanda
  • Takashi Toyama
  • Azusa Shinohara-Kanda
  • Akihiro Iwamatsu
  • Yasuhiro Shinkai
  • Toshiyuki Kaji
  • Makoto Kikushima
  • Yoshito Kumagai
Molecular Toxicology

Abstract

We previously developed a screening method to identify proteins that undergo aggregation through S-mercuration by methylmercury (MeHg) and found that rat arginase I is a target protein for MeHg (Kanda et al. in Arch Toxicol 82:803–808, 2008). In the present study, we characterized another S-mercurated protein from a rat hepatic preparation that has a subunit mass of 42 kDa, thereby facilitating its aggregation. Two-dimensional SDS–polyacrylamide gel electrophoresis and subsequent peptide mass fingerprinting using matrix-assisted laser desorption and ionization time-of-flight mass spectrometry revealed that the 42 kDa protein was NAD-dependent sorbitol dehydrogenase (SDH). With recombinant rat SDH, we found that MeHg is covalently bound to SDH through Cys44, Cys119, Cys129 and Cys164, resulting in the inhibition of its catalytic activity, release of zinc ions and facilitates protein aggregation. Mutation analysis indicated that Cys44, which ligates the active site zinc atom, and Cys129 play a crucial role in the MeHg-mediated aggregation of SDH. Pretreatment with the cofactor NAD, but not NADP or FAD, markedly prevented aggregation of SDH. Such a protective effect of NAD on the aggregation of SDH caused by MeHg is discussed.

Keywords

Methylmercury Covalent modification Cysteine Aggregation Sorbitol dehydrogenase 

Abbreviations

MeHg

Methylmercury

API

Achromobacter protease I

SDH

Sorbitol dehydrogenase

GSH

Glutathione

MALDI-TOF/MS

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

NEM

N-ethylmaleimide

2-ME

2-Mercaptoethanol

MnCl2

Manganese chloride

PVDF

Polyvinylidene difluoride

BPM

Biotin-PEAC5-maleimide

CBB

Coomassie brilliant blue

Notes

Acknowledgments

We thank Dr. Daigo Sumi (Faculty of Pharmaceutical Sciences, Tokushima Bunri University) and Dr. Akira Yasutake (National Institute for Minamata Disease) for helpful advice; Dr. Nobuhiro Shimojo (University of Tsukuba) for his encouragement; Drs. Tomomi Gotoh and Masataka Mori (Department of Molecular Genetics, Graduate School of Medical Sciences, Kumamoto University) for donating the antibody against arginase I. This work was supported by a grant-in-aid (23117703 to Y.K. for scientific research) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Barcelos GR, Grotto D, Serpeloni JM, Angeli JP, Rocha BA, de Oliveira Souza VC, Vicentini JT, Emanuelli T, Bastos JK, Antunes LM, Knasmüller S, Barbosa F Jr (2011) Protective properties of quercetin against DNA damage and oxidative stress induced by methylmercury in rats. Arch Toxicol 85:1151–1157PubMedCrossRefGoogle Scholar
  2. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  3. Clarkson TW (1972) The pharmacology of mercury compounds. Annu Rev Pharmacol 12:375–406PubMedCrossRefGoogle Scholar
  4. Coccini T, Roda E, Castoldi AF, Poli D, Goldoni M, Vettori MV, Mutti A, Manzo L (2011) Developmental exposure to methylmercury and 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB153) affects cerebral dopamine D1-like and D2-like receptors of weanling and pubertal rats. Arch Toxicol 85:1281–1294PubMedCrossRefGoogle Scholar
  5. Hellgren M, Kaiser C, de Haij S, Norberg A, Höög JO (2007) A hydrogen-bonding network in mammalian sorbitol dehydrogenase stabilizes the tetrameric state and is essential for the catalytic power. Cell Mol Life Sci 64:3129–3138PubMedCrossRefGoogle Scholar
  6. Henzel WJ, Billeci TM, Stults JT, Wong SC, Grimley C, Watanabe C (1993) Identifying proteins from two-dimensional gels by molecular mass searching of peptide fragments in protein sequence databases. Proc Natl Acad Sci 90:5011–5015PubMedCrossRefGoogle Scholar
  7. Iwamatsu A, Yoshida-Kubomura N (1996) Systematic peptide fragmentation of polyvinylidene difluoride (PVDF)-immobilized proteins prior to microsequencing. J Biochem 120:29–34PubMedCrossRefGoogle Scholar
  8. Iwamoto N, Nishiyama A, Eiguren-Fernandez A, Hinds W, Kumagai Y, Froines JR, Cho AK, Shinyashiki M (2010) Biochemical and cellular effects of electrophiles present in ambient air samples. Atmos Environ 44:1483–1489CrossRefGoogle Scholar
  9. Jeffery J, Jörnvall H (1988) Sorbitol dehydrogenase. Adv Enzymol Relat Areas Mol Biol 61:47–106PubMedGoogle Scholar
  10. Johansson K, El-Ahmad M, Kaiser C, Jörnvall H, Eklund H, Höög J, Ramaswamy S (2001) Crystal structure of sorbitol dehydrogenase. Chem Biol Interact 130–132:351–358PubMedCrossRefGoogle Scholar
  11. Kanda H, Sumi D, Endo A, Toyama T, Chen CL, Kikushima M, Kumagai Y (2008) Reduction of arginase I activity and manganese levels in the liver during exposure of rats to methylmercury: a possible mechanism. Arch Toxicol 82:803–808PubMedCrossRefGoogle Scholar
  12. Karlsson C, Jörnvall H, Höög JO (1991) Sorbitol dehydrogenase: cDNA coding for the rat enzyme. Variations within the alcohol dehydrogenase family independent of quaternary structure and metal content. Eur J Biochem 198:761–765PubMedCrossRefGoogle Scholar
  13. Kosower NS, Kosower EM (1978) The glutathione status of cells. Int Rev Cytol 54:109–160PubMedCrossRefGoogle Scholar
  14. Maret W, Auld DS (1988) Purification and characterization of human liver sorbitol dehydrogenase. Biochemistry 27:1622–1628PubMedCrossRefGoogle Scholar
  15. Miura T, Shinkai Y, Hirose R, Iwamoto N, Cho AK, Kumagai Y (2011) Glyceraldehyde-3-phosphate dehydrogenase as a quinone reductase in the suppression of 1,2-naphthoquinone protein adduct formation. Free Radic Biol Med 51:2082–2089PubMedCrossRefGoogle Scholar
  16. Nakamura M, Yasutake A, Fujimura M, Hachiya N, Marumoto M (2011) Effect of methylmercury administration on choroid plexus function in rats. Arch Toxicol 85:911–918PubMedCrossRefGoogle Scholar
  17. Rabenstein DL, Fairhurst MT (1975) Nuclear magnetic resonance studies of the solution chemistry of metal complexes. XI. The binding of methylmercury by sulfhydryl-containing amino acids and by glutathione. J Am Chem Soc 97:2086–2092PubMedCrossRefGoogle Scholar
  18. Rasband WS (1997) ImageJ. U.S. National Institutes of Health, Bethesda, Maryland, USA. http://imagej.nih.gov/ij/. Accessed 6 Oct 2006
  19. Rodrigues JL, Serpeloni JM, Batista BL, Souza SS, Barbosa F Jr (2010) Identification and distribution of mercury species in rat tissues following administration of thimerosal or methylmercury. Arch Toxicol 84:891–896PubMedCrossRefGoogle Scholar
  20. Schimke RT (1970) Arginase (rat liver). Methods Enzymol 17A:313–317CrossRefGoogle Scholar
  21. Shinyashiki M, Kumagai Y, Homma-Takeda S, Nagafune J, Takasawa N, Suzuki J, Matsuzaki I, Satoh S, Sagai M, Shimojo N (1996) Selective inhibition of the mouse brain Mn-SOD by methylmercury. Environ Toxicol Pharmacol 2:359–366PubMedCrossRefGoogle Scholar
  22. Shinyashiki M, Kumagai Y, Nakajima H, Nagafune J, Homma-Takeda S, Sagai M, Shimojo N (1998) Differential changes in rat brain nitric oxide synthase in vivo and in vitro by methylmercury. Brain Res 798:147–155PubMedCrossRefGoogle Scholar
  23. Simpson RB (1961) Association constants of methylmercury with sulfhydryl and other bases. J Am Chem Soc 83:4711–4717CrossRefGoogle Scholar
  24. Wagner C, Vargas AP, Roos DH, Morel AF, Farina M, Nogueira CW, Aschner M, Rocha JB (2010) Comparative study of quercetin and its two glycoside derivatives quercitrin and rutin against methylmercury (MeHg)-induced ROS production in rat brain slices. Arch Toxicol 84:89–97PubMedCrossRefGoogle Scholar
  25. Yasutake A, Nakano A, Miyamoto K, Eto K (1997) Chronic effects of methylmercury in rats. I. Biochemical aspects. Tohoku J Exp Med 182:185–196PubMedCrossRefGoogle Scholar
  26. Zou HC, Lü ZR, Wang YJ, Zhang YM, Zou F, Park YD (2009) Effect of cysteine modification on creatine kinase aggregation. Appl Biochem Biotechnol 152:15–28PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Hironori Kanda
    • 1
  • Takashi Toyama
    • 1
  • Azusa Shinohara-Kanda
    • 2
  • Akihiro Iwamatsu
    • 2
    • 5
    • 6
  • Yasuhiro Shinkai
    • 1
  • Toshiyuki Kaji
    • 3
  • Makoto Kikushima
    • 1
  • Yoshito Kumagai
    • 1
    • 4
  1. 1.Doctoral Program in Biomedical Sciences, Graduate School of Comprehensive Human SciencesUniversity of TsukubaTsukubaJapan
  2. 2.Central Laboratories for Key TechnologyKirin Brewery Co., Ltd.YokohamaJapan
  3. 3.Department of Environmental Health, Faculty of Pharmaceutical SciencesTokyo University of ScienceNodaJapan
  4. 4.Faculty of MedicineUniversity of TsukubaTsukubaJapan
  5. 5.Laboratory for Vaccine DesignRIKEN Research Center for Allergy and ImmunologyYokohamaJapan
  6. 6.Protein Research Network, Inc.YokohamaJapan

Personalised recommendations