PMID: 9523703Apr 2, 1998Paper

Characterization of wheat thioredoxin h cDNA and production of an active Triticum aestivum protein in Escherichia coli

European Journal of Biochemistry
M F GautierP Joudrier

Abstract

Two cDNA clones, pTaM13.38 and pTd14.13.2, encoding a Triticum aestivum and a Triticum durum thioredoxin h, respectively, were isolated from mid-maturation seed cDNA libraries. The T aestivum thioredoxin h has a molecular mass of 13.5 kDa and that from T durum has a molecular mass of 13.8 kDa. These two wheat thioredoxin h are 98.5% similar and contain the canonical WCGPC active site and the important structural and functional amino acids that are conserved in thioredoxin sequences. The recombinant T. aestivum thioredoxin h (TrxTa) overproduced in BL21(DE3)pLysS was purified to homogeneity by a three-step procedure including heat treatment, anion-exchange chromatography and gel filtration. TrxTa showed a lower stability to high temperature than Escherichia coli thioredoxin or plant thioredoxin m. The molecular mass of TrxTa, determined by mass spectrometry, is 13,391 Da and corresponds to a protein lacking the first methionine residue, as confirmed by its N-terminal end sequence AASAAT. Using the 5,5'-dithiobis(2-nitrobenzoic acid)-reduction assay and monobromobimane revelation we showed that TrxTa is specifically reduced by wheat NADP:thioredoxin reductase (NTR), and not by E. coli NTR. TrxTa is able to reduce identified targe...Continue Reading

Citations

Dec 6, 2000·European Journal of Biochemistry·J JuttnerU Baumann
Aug 3, 2002·Proceedings of the National Academy of Sciences of the United States of America·Isaac KurekDeborah Delmer
Dec 10, 1999·Proceedings of the National Academy of Sciences of the United States of America·M J ChoB B Buchanan
Mar 21, 2002·Physiologia Plantarum·Eric GelhayeJean-Pierre Jacquot
Oct 11, 2017·Environmental Science and Pollution Research International·Margherita G De BiasiMaurizio Badiani
Sep 13, 2008·The Plant Journal : for Cell and Molecular Biology·Pablo PulidoFrancisco Javier Cejudo
Feb 19, 2011·Expert Review of Proteomics·Gianfranco MamonePasquale Ferranti
Feb 12, 2000·Biochimica Et Biophysica Acta·S D LemaireJ P Jacquot

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