Archives of Pharmacal Research

  • 제24권5호
  • /
  • Pages.446-454
  • /
  • 2001
  • /
  • 0253-6269(pISSN)
  • /
  • 1976-3786(eISSN)
대한약학회 (The Pharmaceutical Society of Korea)
권호 표지

Difference in Susceptibility of Tyrosine Residue to Oxidative Iodination between a Thioredoxin Box Region and a Hormonogenic Region

  • Sok, Dai-Eun (College of Pharmacy, Chungnam National University) ;
  • Charles J.Sih (School of Pharmacy, University of Wisconsin-Madison)
  • 발행 : 2001.10.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Peptide fragments, isolated from proteolytic cleavage of thyroglobulin at specific sites, were examined for the iodination of tyrosine residues. The 50 kDa polypeptide, which was prepared from digestion of bovine thyroglobulin and continuous preparative SDS-PAGE, was subjected to reduction with DTT and alkylation with iodoacetic acid to generate S-car-boxymethylated peptide derivative, which was further hydrohysed by endoproteinase-Asp-N. Peptide products were separated by RP-HPLC, and each fraction was analyzed by LC/ESI-MS and MALDI-MS analyses. Based on the specificity of endoproteinase-Asp-N andthe mass spectra data, a peptide fragment turned out to correspond to a peptide, DALCCVKCPEGSYFQ (1438-1452), characterized by the presence of a thioredoxin box (CVKC) and a tyrosine residue. In addition, another peptide fragment (1453-1465) containing a thioredoxin box (CIPC) and a tyrosine residue was also observed. However, any evidence of iodination of the tyrosine residue present in these peptides was not provided. Meanwhile, tyrosine residues in the peptides, DVEEALAGKYLAGRFA (1366-1381) and DYSGLLLAFQVFLL (1290-1303) were found to be iodinated; mono- or diiodinated tyrosine residues, characteristic of a hormogenic site, existed in both peptides. In addition, the tyrosine residue in the peptide (1218-1252), corresponding to a hormonogenic site was also iodinated. Thus, there was a sharp difference of the susceptibility to oxidative iodination between the tyrosine residue in a hormonogenic site and that in a thioredoxin region. From these results, it is suggested that polypeptide region adjacent to tyrosine residues may govern the susceptibility of tyrosine to oxidative iodination.

키워드

참고문헌

  1. J. Biol. Chem. v.256 Thyroglobulin structurefunction:isolation and characterization of a thyroxinecontaining polypeptide from bovine thyroglobulin Chernoff,S.B.;Rawitch,A.B.
  2. Biochem. Biophys. Res. Comm. v.255 Involvement of oxidative reactions and extracellular protein chaperons in the rescue of misassembled thyroglobulin in the follicular lumen Delom,F.;Lejeune,P.J.;Vinet,L.;Carayon,P.;Mallet,B.
  3. J. Biol. Chem. v.262 The sites of thyroid hormone formation in rabbit thyroglobulin Dunn,J.T.;Anderson,P.C.;Fox,J.W.;Fassler,C.A.;Dunn,A.D.;Hite,L.A.;Moore,R.C.
  4. Endocrinology(3rd Ed.) v.1 Gentile,F.;Di Lauro,R.;Salvatore,G.;DeGroot,L.J.(ed.)
  5. J. Biol. Chem. v.272 Identification of hormonogenic tyrosines in fragment 1218-1591 of bovine throglobulin by mass spectrometry Gentile,F.;Pasquale,F.;Mamone,G.;Malorni,A.;Salvatore,G.
  6. J. Biol. Chem. v.263 A comparison of 30-kDa and 10-kDa hormone-containing fragments of bovine thyroglobulin Gregg,J.D.;Dziadik Turner,C.;Rouse,J.;Hamilton,J.W.;Rawitch,A.D.
  7. J. Biol. Chem. v.381 The thioredoxin boxes of thyroglobulin :possible implications for intermolecular disulfide bond formation in the follicle lumen Klein,Gestmann,U.B.;Schmitz,A.;Herzog,V.
  8. J. Biol. Chem. v.264 Consensus sequences for early iodination and hormonogenesis in human thyroglobulin Lamas,L.;Anderson,P.C.;Fox,J.W.;Dunn,J.T.
  9. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli,U.K.
  10. Free Rad. Res. v.33 ${Cu}^{2+}$-catalyzed oxidative degradation of thyroglobulin Lee,H.J.;Sok,D.E.
  11. Biochem. Biophys. Res. Commun. v.112 Amino acid sequence around a hormonogenic tyrosine residue in the amino terminal region of human thyroglobulin after in vivo and in vitro iodination Marriq,C.;Rolland,M.;Lissitzky,S.
  12. J. Am. Chem. Soc. v.121 Enzymatic mechanism of thyroxine biosynthesis. Identification of the lost three-carbon fragment Ma,Y.A.;Sih,C.J.;Harms,A.
  13. Eur. J. Biochem. v.165 Primary structure of human thyroglobulin deduced from the sequence of its 8448-base complementary DNA Malthiery,Y.;Lissitzky,S.
  14. Nature v.316 Primary structure of bovine thyroglobulin deduced from the sequence of its 8,431-base complementary DNA. Mercken,L.;Simons,M.J.;Swillens,S.;Massaer,M.;Vassart,G.
  15. J. Biol. Chem. v.264 Thyroglobulin-mediated one-and two-electron oxidations of glutathione and ascorbate in thyroid peroxidase systems Nakamura,M.;Yamazaki,I.Kotani,T.;Ohtaki,S.
  16. J. Biol. Chem. v.258 Thyroglobulin structure-function:the amino acid sequence surrounding thyroxine Rawitch,A.B.;Chernoff,S.B.;Litwer,M.R.;Rouse,J.B.;Hamilton,J.W.
  17. Arch. Pharm. Res. v.22 no.5 Regulation and inactivation of brain phosphocholine-phosphatase activity Seo,S.K.;Liu,X.W.;Lee,H.J.;Kim,H.K.;Kim,M.R.;Sok,D.E.
  18. Biochem. Biophys. Acta. v.330 Mechanism of simultaneous iodination and coupling catalyzed by thyroid peroxidase Taurog,A.
  19. Biochimie v.81 The disulfide bond pattern between fragments obtained by the limited proteolysis of bovine thyroglobulin Veneziani,B.M.;Giallauria,F.;Gentile,F.
  20. Comp. Biochem. Physiol. v.122 A comparative review of the structure and biosynthesis of thyroglobulin Venkatesh,S.G.;Deshpande,V.