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cDNA cloning and primary structure analysis of transglutaminase from bluefin tuna Thunnus orientalis

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Abstract

A cDNA clone encoding a tissue-type transglutaminase (TGase) was isolated from the fast muscle of bluefin tuna Thunnus orientalis using a method that combines reverse transcription-PCR and rapid amplification of cDNA ends. The full-length cDNA was 2,872 bp with an open reading frame coding for a protein of 678 amino acid residues. The deduced amino acid sequence had a calculated molecular mass of 74,193 Da and a theoretical isoelectric point of 7.39. The bluefin tuna TGase showed a high amino acid identity of 75% to that of Atlantic salmon Salmo salar, and a low identity of 38% to that of red seabream Pagrus major. Three residues, namely, Cys269, His326, and Asp349, were conserved in a catalytic site, as has also been observed in TGases from other fish and mammals. In phylogenetic analysis, the bluefin tuna TGase cloned in this study was classified into tissue-type TGase2. Southern blot analysis of bluefin tuna genomic DNA digested with restriction enzymes PstI and KpnI using a digoxigenin-labeled probe revealed two bands of different sizes, suggesting the presence of multiple TGase genes in the bluefin tuna.

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References

  1. Folk JE, Finlayson JS (1977) The epsilon-(gamma-glutamyl)lysine crosslink and the catalytic role of transglutaminases. Adv Protein Chem 31:1–133

    Article  PubMed  CAS  Google Scholar 

  2. Folk JE (1980) Transglutaminases. Annu Rev Biochem 49:517–531

    Article  PubMed  CAS  Google Scholar 

  3. Lorand L, Conrad SM (1984) Transglutaminases. Mol Cell Biochem 58:9–35

    Article  PubMed  CAS  Google Scholar 

  4. Greenberg CS, Birckbichler PJ, Rice RH (1991) Transglutaminases: multifunctional cross-linking enzymes that stabilize tissues. FASEB J 5:3071–3077

    PubMed  CAS  Google Scholar 

  5. Joseph D, Lanier TC, Hamann DD (1994) Temperature and pH affect transglutaminase-catalyzed setting of crude fish actomyosin. J Food Sci 59:1018–1023

    Article  CAS  Google Scholar 

  6. Jiang ST, Hsieh JF, Tsai GJ (2004) Interactive effects of microbial transglutaminase and recombinant cystatin on the mackerel and hairtail muscle protein. J Agric Food Chem 52:3617–3625

    Article  PubMed  CAS  Google Scholar 

  7. Yasueda H, Nakanishi K, Kumazawa Y, Nagase K, Motoki M, Matsui H (1995) Tissue-type transglutaminase from red sea bream (Pagrus major). Sequence analysis of the cDNA and functional expression in Escherichia coli. Eur J Biochem 232:411–419

    Article  PubMed  CAS  Google Scholar 

  8. Yeh MS, Liu CH, Hung CW, Cheng W (2009) cDNA cloning, identification, tissue localisation, and transcription profile of a transglutaminase from white shrimp, Litopenaeus vannamei, after infection by Vibrio alginolyticus. Fish Shellfish Immunol 27:748–756

    Article  PubMed  CAS  Google Scholar 

  9. Ikura K, Nasu T, Yokota H, Tsuchiya Y, Sasaki R, Chiba H (1988) Amino acid sequence of guinea pig liver transglutaminase from its cDNA sequence. Biochemistry 27:2898–2905

    Article  PubMed  CAS  Google Scholar 

  10. Ichinose I, Bottenus RE, Davie EW (1990) Structure of transglutaminases. J Biol Chem 265:13411–13414

    PubMed  CAS  Google Scholar 

  11. Griffin M, Casadio R, Bergamini CM (2002) Transglutaminases: nature’s biological glues. Biochem J 368:377–396

    Article  PubMed  CAS  Google Scholar 

  12. Esposito C, Caputo I (2005) Mammalian transglutaminases identification of substrates as a key to physiological function and physiopathological relevance. FEBS J 272:615–631

    Article  PubMed  CAS  Google Scholar 

  13. Nakaoka H, Perez DM, Baek KJ, Das T, Husain A, Misono K, Im MJ, Graham RM (1994) Gh: a GTP-binding protein with transglutaminase activity and receptor signaling function. Science 264:1593–1596

    Article  PubMed  CAS  Google Scholar 

  14. Noguchi K, Ishikawa K, Yokoyama K, Ohtsuka T, Nio N, Suzuki E (2001) Crystal structure of red sea bream transglutaminase. J Biol Chem 276:12055–12059

    Article  PubMed  CAS  Google Scholar 

  15. Sano K, Nakanishi K, Nakamura N, Motoki M, Yasueda H (1996) Cloning and sequence analysis of a cDNA encoding salmon (Onchorhynchus keta) liver transglutaminase. Biosci Biotechnol Biochem 60:1790–1794

    Article  PubMed  CAS  Google Scholar 

  16. Nozawa H, Mamegoshi S, Seki N (1997) Partial purification and characterization of six transglutaminases from ordinary muscles of various fishes and marine invertebrates. Comp Biochem Physiol B Biochem Mol Biol 118:313–317

    Article  PubMed  CAS  Google Scholar 

  17. Itoh Y, Yoshinaka R, Ikeda S (1979) Effects of cysteine and cystine on the gel formation of fish meat by heating. Nippon Suisan Gakkaishi 45:341–345

    Article  CAS  Google Scholar 

  18. Elena SZ, Mahassine A, Rosa O, Evangélica FZ, Casilda N, Juana FL, Esther S, Estrella S, JoséAngel PA (2011) Quality characteristics of dark muscle from yellowfin tuna Thunnus albacares to its potential application in the food industry. Food Nutr Sci 2:22–30

    Article  Google Scholar 

  19. Nakamura YN, Ando M, Seoka M, Kawasaki K, Tsukamasa Y (2007) Changes of proximate and fatty acid compositions of the dorsal and ventral ordinary muscles of the full-cycle cultured Pacific bluefin tuna Thunnus orientalis with the growth. Food Chem 103:234–241

    Article  CAS  Google Scholar 

  20. Albert CM, Campos H, Stampfer MJ, Ridker PM, Manson JE, Willett WC, Ma J (2002) Blood levels of long-chain n-3 fatty acids and the risk of sudden death. N Engl J Med 346:1113–1118

    Article  PubMed  CAS  Google Scholar 

  21. Takahashi M, Minamoto T, Yamashita N, Yazawa K, Sugimura T, Esumi H (1993) Reduction in formation and growth of 1,2-dimethylhydrazine-induced aberrant crypt foci in rat colon by docosahexaenoic acid. Cancer Res 53:2786–2789

    PubMed  CAS  Google Scholar 

  22. Zhang J, Sasaki S, Amano K, Kesteloot H (1999) Fish consumption and mortality from all causes, ischemic heart disease, and stroke: an ecological study. Prev Med 28:520–529

    Article  PubMed  CAS  Google Scholar 

  23. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  PubMed  CAS  Google Scholar 

  24. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTALW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  PubMed  CAS  Google Scholar 

  25. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    PubMed  CAS  Google Scholar 

  26. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  PubMed  CAS  Google Scholar 

  27. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1987) Current protocols in molecular biology. Green Publishing Associates and Wiley-Interscience, New York

    Google Scholar 

  28. Ichinose A, Hendrickson LE, Fujikawa K, Davie EW (1986) Amino acid sequence of the a subunit of human factor XIII. Biochemistry 25:6900–6906

    Article  PubMed  CAS  Google Scholar 

  29. Molhuizen HO, Alkemade HA, Zeeuwen PL, de Jongh GJ, Wieringa B, Schalkwijk J (1993) SKALP/elafin: an elastase inhibitor from cultured human keratinocytes. Purification, cDNA sequence, and evidence for transglutaminase cross-linking. J Biol Chem 268:12028–12032

    PubMed  CAS  Google Scholar 

  30. Kim IG, Gorman JJ, Park SC, Chung SI, Steinert PM (1993) The deduced sequence of the novel protransglutaminase E (TGase3) of human and mouse. J Biol Chem 268:12682–12690

    PubMed  CAS  Google Scholar 

  31. Király R, Csosz E, Kurtán T, Antus S, Szigeti K, Simon-Vecsei Z, Korponay-Szabó IR, Keresztessy Z, Fésüs L (2009) Functional significance of five noncanonical Ca2+-binding sites of human transglutaminase 2 characterized by site-directed mutagenesis. FEBS J 276:7083–7096

    Article  PubMed  Google Scholar 

  32. Begg GE, Holman SR, Stokes PH, Matthews JM, Graham RM, Iismaa SE (2006) Mutation of a critical arginine in the GTP-binding site of transglutaminase 2 disinhibits intracellular cross-linking activity. J Biol Chem 281:12603–12609

    Article  PubMed  CAS  Google Scholar 

  33. Yee VC, Le Trong I, Bishop PD, Pedersen LC, Stenkamp RE, Teller DC (1996) Structure and function studies of factor XIIIa by X-ray crystallography. Semin Thromb Hemost 22:377–384

    Article  PubMed  CAS  Google Scholar 

  34. Yee VC, Pedersen LC, Le Trong I, Bishop PD, Stenkamp RE, Teller DC (1994) Three-dimensional structure of a transglutaminase: human blood coagulation factor XIII. Proc Natl Acad Sci USA 91:7296–7300

    Article  PubMed  CAS  Google Scholar 

  35. Pedersen LC, Yee VC, Bishop PD, Le Trong I, Teller DC, Stenkamp RE (1994) Transglutaminase factor XIII uses proteinase-like catalytic triad to crosslink macromolecules. Prot Sci 3:1131–1135

    Article  CAS  Google Scholar 

  36. Han BG, Cho JW, Cho YD, Jeong KC, Kim SY, Lee BI (2010) Crystal structure of human transglutaminase 2 in complex with adenosine triphosphate. Int J Biol Macromol 47:190–195

    Article  PubMed  CAS  Google Scholar 

  37. Deasey S, Grichenko O, Du S, Nurminskaya M (2012) Characterization of the transglutaminase gene family in zebrafish and in vivo analysis of transglutaminase-dependent bone mineralization. Amino Acids 42:1065–1075

    Article  PubMed  CAS  Google Scholar 

  38. Sugitani K, Matsukawa T, Maeda A, Kato S (2006) Upregulation of transglutaminase in the goldfish retina during optic nerve regeneration. Adv Exp Med Biol 572:525–530

    Article  PubMed  CAS  Google Scholar 

  39. Koh D, Inohaya K, Imai Y, Kudo A (2004) The novel medaka transglutaminase gene is expressed in developing yolk veins. Gene Expr Patterns 4:263–266

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Laboratory of Marine Biochemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo, 113-8657, Japan

    Koki Ikeguchi, Gen Kaneko & Shugo Watabe

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  1. Koki Ikeguchi
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  2. Gen Kaneko
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  3. Shugo Watabe
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Correspondence to Shugo Watabe.

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Ikeguchi, K., Kaneko, G. & Watabe, S. cDNA cloning and primary structure analysis of transglutaminase from bluefin tuna Thunnus orientalis . Fish Sci 78, 667–674 (2012). https://doi.org/10.1007/s12562-012-0474-8

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  • DOI: https://doi.org/10.1007/s12562-012-0474-8

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