PMID: 11903963Mar 21, 2002Paper

Isolation and characterization of an extended thioredoxin h from poplar

Physiologia Plantarum
Eric GelhayeJean-Pierre Jacquot

Abstract

A cDNA coding for a thioredoxin h has been isolated from a xylem/phloem poplar cDNA library by RACE-PCR. The nucleotide sequence called popTrx-h2 is homologous to other thioredoxins h isolated from plants but differs from the other thioredoxins h by presenting a 30 amino acid long N-terminus extension. A variant of this cDNA lacking the N-terminal extension was also generated by PCR. Both cDNAs have been introduced into an expression plasmid (pET-3d) and the recombinant proteins have been expressed to a high level and purified from Escherichia coli cells. Protein sequencing showed that a part of the N-terminal extension was cleaved in the E. coli cells, with the first 19 amino acids missing, suggesting the presence of a putative cleavage site in the N-terminal extension of popTrx-h2. Both recombinant proteins display unusual catalytic properties compared to other thioredoxins h characterized so far, i.e. a weak reduction by Arabidopsis thaliana NADPH-dependent thioredoxin reductase, and a weak activation of the chloroplastic NADP-malate dehydrogenase, a non-physiological target enzyme. Northern blot experiments indicate that the transcripts of popTrx-h2 are present in leaves and roots, albeit at a lower level compared to the ea...Continue Reading

References

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Mar 5, 1990·Journal of Molecular Biology·S K KattiH Eklund
Dec 5, 1968·European Journal of Biochemistry·A Holmgren
Jan 1, 1995·Methods in Enzymology·J P JacquotM Miginiac-Maslow
Jun 6, 1995·Proceedings of the National Academy of Sciences of the United States of America·R Rivera-MadridY Meyer
Aug 26, 1998·Biochemical and Biophysical Research Communications·Y Huang, F E Domann
Jul 3, 1999·The Journal of Biological Chemistry·L VerdoucqY Meyer
Feb 12, 2000·Biochimica Et Biophysica Acta·S D LemaireJ P Jacquot
Aug 31, 2000·Journal of Molecular Biology·G CapitaniP Schürmann
Mar 12, 2004·Annual Review of Plant Physiology and Plant Molecular Biology·P. Schurmann, J.-P. Jacquot
Aug 1, 1997·The New Phytologist·Jean-Pierre JacquotYves Meyer

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Citations

Oct 13, 2006·Photosynthesis Research·Eric GelhayeJean-Pierre Jacquot
Sep 24, 2004·Proceedings of the National Academy of Sciences of the United States of America·Eric GelhayeJean-Pierre Jacquot
Mar 10, 2011·FEBS Letters·Kamel ChibaniNicolas Rouhier
May 4, 2004·Plant Physiology and Biochemistry : PPB·Eric GelhayeJean-Pierre Jacquot
Dec 17, 2003·FEBS Letters·Eric GelhayeJean Pierre Jacquot

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