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Multimerization of Bovine Thyroglobulin, Partially Unfolded or Partially Unfolded/Reduced; Involvement of Protein Disulfide Isomerase and Glutathionylated Disulfide Linkage

Liu, Xi-Wen    (College of Pharmacy, Chungnam National University   ); Sok , Dai-Eun    (College of Pharmacy, Chungnam National University  );
  • 초록

    Fate of the nascent thyrolglobulin (Tg) molecule is characterized by multimerization. To establish the formation of Tg multimers, the partially unfolded/reduced Tg or deoxycholate-treated/ reduced Tg was subjected to protein disulfide isomerase (PDI)-mediated multimerization. Oxidized glutathione/PDI-mediated formation of multimeric Tg forms, requiring at least an equivalent molar ratio of PDI/Tg monomer, decreased with increasing concentration of reduced glutathione (GSH), suggesting the oxidizing role of PDI. Additional support was obtained when PDI alone, at a PDI/Tg molar ratio of 0.3, expressed a rapid multimerization. Independently, the exposure of partially unfolded Tg to GSH resulted in Tg multimerization, enhanced by PDI, according to thiol-disulfide exchange. Though to a lower extent, a similar result was observed with the dimerization of deoxycholate-pretreated Tg monomer. Consequently, it is implied that intermolecular disulfide linkage may be facilitated at a limited region of unfolded Tg. In an attempt to examine the multimerization site, the cysteine residue-rich fragments of the Tg were subjected to GSH-induced multimerization; a 50 kDa fragment, containing three vicinal dithiols, was multimerized, while an N-terminal domain was not. Present results suggest that the oxidase as well as isomerase function of PDI may be involved in the multimerization of partially unfolded Tg or deoxycholate-treated Tg.


  • 주제어

    Multimerization .   Thyroglobulin (Tg) .   Partially unfolded reduced .   Protein disulfide isomerase(PDI) .   Disulfide.  

  • 참고문헌 (29)

    1. Freedman, R. B., Trends Biochem. Sci., 9, 438-441 (1984) 
    2. Nigam, S. K., Goldberg, A. L., Ho, S., Rohde, M. F., Bush, K. T., and Sherman, M. Y., A set of endoplasmic reticulum proteins possessing properties of molecular chaperones includes Ca(2+)-binding proteins and members of the thioredoxin superfamily. J. BioI. Chem., 269, 1744-1749 (1994) 
    3. Ellman, G. L., Tissue sulfhydryl groups. Arch. Biochem. Biophys., 82, 70-77 (1959) 
    4. Gentile, F., Di Lauro, R., and Salvatore, G., in Endocrinology, In DeGroot, L. J., 3rd Ed., pp. 517-542, W. B. Saunders Co., Philadelphia, PA (1995) 
    5. Gilbert, H. F., Molecular and cellular aspects of thiol-disulfide exchange. Adv. Enzymol. Relat. Areas Mol. BioI., 63, 69-172 (1990) 
    6. Klein, M., Gestmann, I., Berndorfer, U., Schmitz, A., and Herzog v., The thioredoxin boxes of thyroglobulin: possible implications for intermolecular disulfide bond formation in the follicle lumen. BioI. Chem., 381, 593-601 (2000) 
    7. Delom, F., Mallet, B., Carayon, P, and Lejeune, P J., Role of extracellular molecular chaperones in the folding of oxidized proteins. Refolding of colloidal thyroglobulin by protein disulfide isomerase and immunoglobulin heavy chain-binding protein. J. BioI. Chem., 276, 21337-21342 (2001) 
    8. Gentile, F., Pasquale, F., Mamone, G., Malorni, A., and Salvatore, G., Identification of Hormonogenic Tyrosines in Fragment 1218-1591 of Bovine Thyroglobulin by Mass Spectrometry. Hormonogenic acceptor Tyr-1291 and donor Tyr-1375. J. BioI. Chem., 272, 639-646 (1997) 
    9. Gilbert, H. F., Catalysis of thiol/disulfide exchange: singleturnover reduction of protein disulfide-isomerase by glutathione and catalysis of peptide disulfide reduction. Biochemistry, 28, 7298-7305 (1989) 
    10. Kim, P. S., Kim, K.-R., and Arvan, P., Disulfide-linked aggregation of thyroglobulin normally occurs during nascent protein folding. Am. J. Physiol. Cell Physiol., 265, C704-C711 (1993) 
    11. De Crombrugghe, B., Pitt-Rivers, R., and Edelhoch, H., The Properties of Thyroglobulin. XI. The reduction of the disulfide bonds. J. BioI. Chem., 241, 2766-2773 (1966) 
    12. Delom, F., Lejeune, P.-J., Vinet, L., Carayon, P., and Mallet, B., Involvement of oxidative reactions and extracellular protein chaperones in the rescue of misassembled thyroglobulin in the follicular lumen. Biochem. Biophys. Res. Comm., 255, 438-443 (1999) 
    13. Malthiery, Y. and Lissitzky, S., Primary structure of human thyroglobulin educed from the sequence of its 8448-base complementary DNA Eur. J. Biochem., 165, 491-498 (1987) 
    14. Frand, A. R., Cuozzo, J. W., and Kaiser, C. A, Pathways for protein disulphide bond formation. Trends Cell BioI., 10, 203-210 (2000) 
    15. Chernoff, S. B. and Rawitch, A .B., Thyroglobulin structurefunction. Isolation and characterization of a thyroxinecontaining polypeptide from bovine thyroglobulin. J. BioI. Chem., 256, 9425-9430 (1981) 
    16. Kim, P. S. and Arvan, P., Calnexin and BiP act as sequential molecular chaperones during thyroglobUlin folding in the endoplasmic reticulum. J. Cell BioI., 128, 29-38 (1995) 
    17. Herzog, V., Berndorfer, U., and Saber, Y., Isolation of insoluble secretory product from bovine thyroid: extracellular storage. of thyroglobulin in covalently cross-linked form. J. Cell BioI., 118, 1071-1083 (1992) 
    18. Pagani, M., Fabbri, M., Benedetti, C., Fassio, A., Pilati, S., Bulleid, N. J., Cabibbo, A, and Sitia, R., Endoplasmic Reticulum Oxidoreductin 1-L$\beta$ (ER01-L$\beta$), a Human Gene Induced in the Course of the Unfolded Protein Response. J. BioI. Chem., 275, 23685-23692 (2000) 
    19. Mercken, L., Simons, M.-J., Swillens, S., Massaer, M., and Vassart, G., Primary structure of bovine thyroglobulin deduced from the sequence of its 8,431-base complementary DNA. Nature, 316, 647-651 (1985) 
    20. Freedman, R. B., Hawkins, H. C., and McLaughlin, S. H., Protein disulfide-isomerase. Methods Enzymol., 251, 397-406 (1995) 
    21. Ruoppolo, M., Freedman, R. B., Pucci, P., and Marino, G., Glutathione-dependent pathways of refolding of RNase T1 by oxidation and disulfide isomerization: catalysis by protein disulfide isomerase. Biochemistry, 35,13636-13646 (1996) 
    22. Bass, R., Ruddock, L. W., Klappa, P., and freedman, R. B., A major fractin of endoplasmic reticulum-located glutathione is present as mixed disulfides with protein. J. Biol. Chem., 279, 5257-5262 (2004) 
    23. Cuozzo, J. W. and Kaiser, C. A, Competition between glutathione and protein thiols for disulphide-bond formation. Nature Cell BioI., 1, 130-135 (1999) 
    24. Kuznetsov, G., Chen, L. B., and Nigam, S. K., Multiple Molecular Chaperones Complex with Misfolded Large Oligomeric Glycoproteins in the Endoplasmic Reticulum. J. BioI. Chem., 272,3057-3063 (1997) 
    25. Freedman, R. B., Hirst, T. R., and Tuite, M. F., Protein disulphide isomerase: building bridges in protein folding. Trends Biochem. Sci., 19, 331-336 (1994) 
    26. Laemmli, U. K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685 (1970) 
    27. Darby, N. J., Freedman, R. B., and Creghton, T. E., Dissecting the mechanism of protein disulfide isomerase: catalysis of disulfide bond formation in a model peptide. Biochemistry, 33,7937-7947 (1994) 
    28. Gilbert, H. F., Protein Disulfide Isomerase and Assisted Protein Folding. J. BioI. Chem., 272, 29399-29402 (1997) 
    29. Bradford, M. M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72, 248-254 (1976) 

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