Purification of Carp Serum Metallothionein Induced in Carp with Cadmium and Production of Polyclonal Antibody

카드뮴 유도 잉어 혈액으로부터의 Metallothionein 정제 및 이에 대한 다클론 항체 생산

  • Received : 2009.10.13
  • Accepted : 2009.11.16
  • Published : 2009.11.30

Abstract

Metallothioneins (MTs) are low molecular weight, cysteine-rich, and heavy metal binding proteins, which could be induced with heavy metals such as Cd, Hg, Zn and Cu in liver, kidney, and in cultured cells. By using ion exchange chromatography on DE-52, MT was purified from the serum of carp induced with cadmium in order to produce antibody against MT. Polyclonal antibody produced against purified carp MT reacted well with MT in the serum of carp induced with cadmium, whereas control serum did not. This may indicate that the polyclonal antibody against the carp MT could be used for the preparation of biosensors to detect MT in fishes like carp.

Keywords

Acknowledgement

Supported by : 한국식품연구원

References

  1. Brower M, Enghild J, Holxum-Browwer T, Thogersen I, TruncaliA. 1995. Primary structure and tissue-specific expression of bluecrab (Callinectes sapidus) metallothionein isoforms. Biochem. J.311: 617-622
  2. Campenhour KV, Infante HG, Adams F, Blust R. 2004. Inductionand binding of Cd, Cu, and Zn to metallothionein in carp (Cyprinuscarpio) using HPLC-ICP-TOFMS. Toxicol. Sci. 80: 276-287 https://doi.org/10.1093/toxsci/kfh149
  3. Chatterjee A, Maiti IB. 1987. Purification and immunologicalcharacterization of catfish (Heteropneustes fossilis) metallothionein.Mol. Cell. Biochem. 78: 55-64
  4. Cousins RJ. 1985. Absorption, transports and hepatic metabolismof copper and zinc : special reference to metallothionein andceruloplasmin. Physiol. Rev. 65: 238-309
  5. Dunn M, Blalock TL, Cousins RJ. 1987. Metallothionein. ProcSoc Exp. Biol. Med. 185: 107-119
  6. Eaton, DL, Toal, BF. 1982. Evaluation of the Cd/hemoglobinaffinity assay for the rapid determination of metallothionein inbiological tissues. Toxicol. Appl. Pharmacol. 66: 134-142 https://doi.org/10.1016/0041-008X(82)90068-0
  7. Gao D, Wnag GT, Chen XT, Nie P. 2009. Metallothionein-2 genefrom the mandarin fish Siniperca chuasti: cDNA cloning, tissueexpression, and immunohistochemical localization. Comp. Biochem.Physiol. 149C: 18-25
  8. George SG, Todd K, Wright J. 1996. Regulation of metallothioneinin teleosts: induction of MT mRNA and protein by cadmiumin hepatic and extra hepatic tissue of a marine flat fishthe turbot (Scophthalmus maximus). Comp. Biochem. Physiol.113C: 109-115
  9. Gopal V, Parvathy S, Balassubramanian PR. 1997. Effect of heavymetals on the blood protein biochemistry of the fish Cyprinuscarpio and its use as a bio-indicator of pollution stress. Environ.Monitor. Assess. 48: 117-124 https://doi.org/10.1023/A:1005767517819
  10. Hamer DH, Thieles DJ, Lemantt JF. 1985. Function and autoregulationof yeast copperthionein. Science 228: 685-690 https://doi.org/10.1126/science.3887570
  11. Järup L. 2003. Hazards of heavy metal contamination. Br. Med.Bull. 68: 167-182 https://doi.org/10.1093/bmb/ldg032
  12. Jensen LT, Howare WR, Strain JJ, Winge DR, Culotta VC. 1996.Enhanced effectiveness of copper ion buffering by Cup1 metallothioneincompared with CRS5 metallothionein in Saccharomycescerevisiae. J. Biol. Chem. 271: 18514-18519 https://doi.org/10.1074/jbc.271.31.18514
  13. Kägi JHR, Kojima Y. 1987. Chemistry and biochemistry of metallothionein.Exp. Suppl. 52: 25-61
  14. Kim N, Kim D-K, Cho Y-J, Moon D-K, Kim W-Y. 2008. Carpvitellogenin detection by an optical waveguide lightmode spectroscopybiosensor. Biosens. Bioelectron. 24: 391-396 https://doi.org/10.1016/j.bios.2008.04.013
  15. Laemmli, UK.1970. Cleavage of structural proteins during assemblyof the head of bacteriophage T4. Nature. 227: 680-685 https://doi.org/10.1038/227680a0
  16. Margoshes M, Vales BL. 1957. A cadmium protein from equinekidney cortex. J. Am. Chem. Soc. 79: 4813-4814 https://doi.org/10.1021/ja01574a064
  17. Mance G. 1987. Pollution threat of heavy metals in aquatic environments.Elsevier, London, UK. p. 372
  18. Moon D-K, Kim N, Kim WY. 2006. Reactivity of the antibodiesagainst purified carp vitellogenin and a synthetic vitellogeninpeptide. J. Korean Soc. Appl. Biol. Chem. 49: 196-201
  19. Olafson PW, McCubbin WD, Kay MC. 1988. Primary and secondarystructural analysis of a unique prokaryotic metallothioneinfrom Synechococcus sp cyanobacterium. Bio. Chem. J.251: 691-699
  20. Pan A, Wang Z, Ru B. 1991. Determination and purification ofmetallothionein by high performance liquid chromatography.Biomed. Chromatogr. 5: 193-197 https://doi.org/10.1002/bmc.1130050503
  21. Pan A, Tie F, Duan Z, Ma H, Li L, Ru B. 1993. Purification andcharacterization of hedgehog liver metallothionein. Biomed.Chromatogr. 7: 94-98 https://doi.org/10.1002/bmc.1130070210
  22. Price NM, Morel FMM. 1995. Cadmium and cobalt substitutionfor zinc in a marine diatom. Nature 344: 658-660 https://doi.org/10.1038/344658a0
  23. Roesijadi G. 1992. Metallothioneins in metal regulation and toxicityin aquatic animals. Aqua. Toxicol . 22: 81-114 https://doi.org/10.1016/0166-445X(92)90026-J
  24. Sampath-Kunar R, Munro AD, Lam TJ. 1996. Ontogeny ofimmunoreactive steroidogenic proteins (Adrenodoxin and Cytochromep-450) in the interrenals of the teleost Lates calcarifer.Gen. Comp. Endocrine. 102: 147-155 https://doi.org/10.1006/gcen.1996.0056
  25. Scheuhammer AM, Cherian MG. 1986. Quantification of metallothioneinby a silver-saturation method. Toxicol. Appl. Pharmacol. 82: 417-425 https://doi.org/10.1016/0041-008X(86)90277-2
  26. Syring RA, Hoexum B, Brouwer M. 2000. Cloning and sequencingof cDNA encoding for a novel copper-specific metallothioneinand two cadmium-inducible metallothioneins from the bluecrab Callinectes sapidus. Comp. Biochem. Physiol. 125: 325-332 https://doi.org/10.1016/S1095-6433(00)00157-4
  27. Turánek J, Kováø J, Zábrššová, H. 1987. Rapid purification of theisoforms of rat liver Cd, Zn-metallothionein by high-performanceliquid chromatography. J. Chromatogr. 410: 470-473 https://doi.org/10.1016/S0021-9673(00)90079-7
  28. Vosyliene MZ, Jankaite A. 2006. Effect of heavy metal modelmixture on rainbow trout biological parameters. Ekologija. 4:12-17
  29. Witeska M. 2005. Stress in fish-hematological and immunologicaleffects of heavy metals. Electron. J. Ichthyol. 1: 35-41
  30. Wu SM, Weng CF, Yu MJ, Lin CC, Chen ST, Hwang JC, HwangPP. 1999. Cadmium-inducible metallothionein in Tilapia (Oreochromismossambicus). Bull. Environ. Contam. Toxicol. 62:758-768 https://doi.org/10.1007/s001289900937
  31. Yadv N, Khandelwal S. 2005. Effect of picroliv on cadmiuminducedhepatic and renal damage in the rat. Hum. Exp. Toxicol.25: 581-591 https://doi.org/10.1177/096032706072455
  32. Yeoh CG, Schreck CB, Fitzpatrick MS, Feist GW. 1996. In vivosteroid metabolism in embryonic and newly hatched trout(Orcorhynchusmykiss). Gen. Comp. Endocrinol. 102: 197-209 https://doi.org/10.1006/gcen.1996.0061