NMR Studies of the Interaction of Tryparedoxin with Redox-Inactive Substrate Homologues
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- 作者：Dirk Krumme ; Heike Budde ; Hans-Jü ; rgen Hecht ; Ulrich Menge ; Oliver Ohlenschlä ; ger ; Anton Ross ; Josef Wissing ; Victor Wray ; and Leopold Flohé
- 刊名：Biochemistry
- 出版年：2003
- 出版时间：December 23, 2003
- 年：2003
- 卷：42
- 期：50
- 页码：14720 - 14728
- 全文大小：725K
- 年卷期：v.42,no.50(December 23, 2003)
- ISSN：1520-4995
文摘
Tryparedoxins (TXNs) are trypanothione-dependent peroxiredoxin oxidoreductases involvedin hydroperoxide detoxification that have been shown to determine virulence in trypanosomatids. Thestructure of 15N,13C-doubly-labeled, C-terminally-His-tagged tryparedoxin 1 from Crithidia fasciculata(Cf TXN1) was elucidated by three-dimensional NMR spectroscopy. Global folding was found to besimilar to the crystal structure, but regions near the active site, especially the onset of helix 
chars/alpha.gif" BORDER=0>1 with theredox-active Cys 43 and helix chars/alpha.gif" BORDER=0>2 relevant to substrate binding, were less well defined in solution. Theredox-inactive inhibitory substrate analogue N1,N8-bis(ophthalmyl)spermidine was used to study thesubstrate/TXN interaction by two-dimensional 1H,15N NMR spectroscopy. The NMR data complementedby molecular modeling revealed several alternative modes of ligand binding. The results confirm andextend the concept of TXN action and specificity derived from X-ray analysis and site-directed mutagenesisand thus improve the rational basis for inhibitor design.
chars/alpha.gif" BORDER=0>1 with theredox-active Cys 43 and helix chars/alpha.gif" BORDER=0>2 relevant to substrate binding, were less well defined in solution. Theredox-inactive inhibitory substrate analogue N1,N8-bis(ophthalmyl)spermidine was used to study thesubstrate/TXN interaction by two-dimensional 1H,15N NMR spectroscopy. The NMR data complementedby molecular modeling revealed several alternative modes of ligand binding. The results confirm andextend the concept of TXN action and specificity derived from X-ray analysis and site-directed mutagenesisand thus improve the rational basis for inhibitor design.