Biochemical characterization of thioredoxin 1 from <i>Dictyostelium discoideum</i>

Wetterauer, Birgit; Veron, M; Miginiac‐Maslow, Myroslawa; Decottignies, Paulette; Jacquot, Jean‐Pierre

doi:10.1111/j.1432-1033.1992.tb17331.x

Cited by 20 publications

(19 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…FBPase activation was performed by incubating DTT-reduced Trx with 2 M FBPase. The enzyme activity was measured on aliquots by a coupled assay (28). Prx activity was assayed in the presence of a 20 M concentration of a 2-Cys Prx from Arabidopsis, 400 M DTT, 400 M t-BOOH, and variable amounts of Trx.…”

Section: Methodsmentioning

confidence: 99%

The Arabidopsis Plastidial Thioredoxins

Collin

Issakidis‐Bourguet²,

Marchand³

et al. 2003

Journal of Biological Chemistry

Self Cite

314

105

View full text Add to dashboard Cite

The sequencing of the genome of Arabidopsis thaliana revealed that this plant contained numerous isoforms of thioredoxin (Trx), a protein involved in thiol-disulfide exchanges. On the basis of sequence comparison, seven putative chloroplastic Trxs have been identified, four belonging to the m-type, two belonging to the f-type, and one belonging to a new x-type. In the present work, these isoforms were produced and purified as recombinant proteins without their putative transit peptides. Their activities were tested with two known chloroplast thioredoxin targets: NADP-malate dehydrogenase and fructose-1,6-bisphosphatase and also with a chloroplastic 2-Cys peroxiredoxin. The study confirms the strict specificity of fructose-bisphosphatase for Trx f, reveals that some Trxs are unable to activate NADP-malate dehydrogenase, and shows that the new x-type is the most efficient substrate for peroxiredoxin while being inactive toward the two other targets. This suggests that this isoform might be specifically involved in resistance against oxidative stress. Three-dimensional modeling shows that one of the m-type Trxs, Trx m3, which has no activity with any of the three targets, exhibits a negatively charged surface surrounding the active site. A green fluorescent protein approach confirms the plastidial localization of these Trxs.

show abstract

Section: Methodsmentioning

confidence: 99%

The Arabidopsis Plastidial Thioredoxins

Collin

Issakidis‐Bourguet²,

Marchand³

et al. 2003

Journal of Biological Chemistry

Self Cite

314

105

View full text Add to dashboard Cite

show abstract

“…The oxidized form of thioredoxin, having an intramolecular disulfide bridge between the two cysteine residues from the catalytic center, is generally reduced by a thioredoxin reductase. Thioredoxins are ubiquitous and have been described in prokaryotes as well as in eukaryotes, including, fungi, plants, invertebrates, and vertebrates (2)(3)(4). Higher plants are known to possess at least three types of thioredoxins: thioredoxin f and m, which are both encoded by the nuclear genome but are located within the chloroplast, and thioredoxin h, which is a cytosolic protein.…”

mentioning

confidence: 99%

In vivo functional discrimination between plant thioredoxins by heterologous expression in the yeast Saccharomyces cerevisiae

Mouaheb

Thomas

Verdoucq

et al. 1998

Proc. Natl. Acad. Sci. U.S.A.

113

View full text Add to dashboard Cite

Whereas vertebrates possess only two thioredoxin genes, higher plants present a much greater diversity of thioredoxins. For example, Arabidopsis thaliana has five cytoplasmic thioredoxins (type h) and at least as many chloroplastic thioredoxins. The abundance of plant thioredoxins leads to the question whether the various plant thioredoxins play a similar role or have specific functions. Because most of these proteins display very similar activities on artificial or biological substrates in vitro, we developed an in vivo approach to answer this question. The disruption of both of the two Saccharomyces cerevisiae thioredoxin genes leads to pleiotropic effects including methionine auxotrophy, H 2 O 2 hypersensitivity, altered cell cycle characteristics, and a limited ability to use methionine sulfoxide as source of methionine. We expressed eight plant thioredoxins (six cytoplasmic and two chloroplastic) in yeast trx1, trx2 double mutant cells and analyzed the different phenotypes. Arabidopsis type h thioredoxin 2 efficiently restored sulfate assimilation whereas Arabidopsis type h thioredoxin 3 conferred H 2 O 2 tolerance. All thioredoxins tested could complement for reduction of methionine sulfoxide, whereas only type h thioredoxins were able to complement the cell cycle defect. These findings clearly indicate that specific interactions between plant thioredoxins and their targets occur in vivo.Thioredoxins are small oxidoreductases (molecular mass Ϸ12 kDa) that all contain two redox-active half-cystine residues in an exposed active center, having the amino acid sequence WCXPC (1). In its reduced form, thioredoxin can function as hydrogen donor for a variety of target proteins. The oxidized form of thioredoxin, having an intramolecular disulfide bridge between the two cysteine residues from the catalytic center, is generally reduced by a thioredoxin reductase. Thioredoxins are ubiquitous and have been described in prokaryotes as well as in eukaryotes, including, fungi, plants, invertebrates, and vertebrates (2-4). Higher plants are known to possess at least three types of thioredoxins: thioredoxin f and m, which are both encoded by the nuclear genome but are located within the chloroplast, and thioredoxin h, which is a cytosolic protein.The chloroplastic thioredoxins f and m are thought to provide a functional link between the light-absorbing pigments and several key metabolic enzymes such as fructose-1,6-bisphosphatase, phosphoribulose kinase, and malate dehydrogenase (5-7). Both oxidized thioredoxin f and thioredoxin m are reduced by a ferredoxin-dependent thioredoxin reductase (8-10). In contrast, the cytosolic thioredoxins h are reduced by an NADPH-dependent thioredoxin reductase (11,12). Recent progress in the systematic sequencing of plant genomes provides evidence that the diversity of thioredoxins is much greater than previously expected, each class of thioredoxin being encoded by multigene families. For instance, Arabidopsis thaliana has been shown to express at least two thioredoxins f (L.V. and...

show abstract

“…In vivo, thioredoxins are reduced by a thioredoxin reductase. Thioredoxins have been isolated from prokaryotes and eukaryotes, including vertebrates, invertebrates, and fungi (2,3,41). Higher plants present two independent thioredoxin systems: the chloroplastic system consists of two nuclear gene-encoded proteins, thioredoxins m and f, which are reduced by a ferredoxindependent thioredoxin reductase (4,5), and regulates the dark/light-related cycles (6,7).…”

mentioning

confidence: 99%

Evidence for five divergent thioredoxin h sequences in Arabidopsis thaliana.

Rivera‐Madrid

Mestres

Marinho

et al. 1995

Proc. Natl. Acad. Sci. U.S.A.

111

View full text Add to dashboard Cite

Five different clones encoding thioredoxin homologues were isolated from Arabidopsis thaliana cDNA libraries. On the basis of the sequences they encode divergent proteins, but all belong to the cytoplasmic thioredoxins h previously described in higher plants. The five proteins obtained by overexpressing the coding sequences in Escherichia coli present typical thioredoxin activities (NADP+-malate dehydrogenase activation and reduction by Arabidopsis thioredoxin reductase) despite the presence of a variant active site, Trp-Cys-Pro-Pro-Cys, in three proteins in place of the canonical Trp-Cys-Gly-Pro-Cys sequence described for thioredoxins in prokaryotes and eukaryotes. Southern blots show that each cDNA is encoded by a single gene but suggest the presence of additional related sequences in theArabidopsis genome. This very complex diversity of thioredoxins h is probably common to all higher plants, since the Arabidopsis sequences appear to have diverged very early, at the beginning of plant speciation. This diversity allows the transduction of a redox signal into multiple pathways.Thioredoxins are small proteins (105-120 aa) that contain the strictly conserved sequence Trp-Cys-Gly-Pro-Cys (1). The reduced cysteine pair forms a very reactive center able to disrupt the disulfide bridge of target proteins. In vivo, thioredoxins are reduced by a thioredoxin reductase. Thioredoxins have been isolated from prokaryotes and eukaryotes, including vertebrates, invertebrates, and fungi (2, 3, 41). Higher plants present two independent thioredoxin systems: the chloroplastic system consists of two nuclear gene-encoded proteins, thioredoxins m and f, which are reduced by a ferredoxindependent thioredoxin reductase (4, 5), and regulates the dark/light-related cycles (6, 7). In addition, the presence of a thioredoxin h system that is reduced by an NADP-dependent thioredoxin reductase was demonstrated some years ago in heterotrophic plant tissues (8, 9), but the first sequences for plant thioredoxins h (10-13) and NADP-dependent thioredoxin reductase (14) were obtained only recently. In this paper, we show that the Arabidopsis thaliana genome encodes at least five very divergent thioredoxin h genes, § a situation similar to the one of Dictyostelium discoideum (15) The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. Toulouse) were subcultured every 2 weeks by 10-fold dilution in B5 medium (42) with sucrose (20 mg/ml) and 2,4-dichlorophenoxyacetic acid (1 ,ug/ml) and collected after 3 days of subculture (growth phase). Primary calli (seeds germinated on the callus medium) and established calli (the cell suspension grown on the callus medium) were grown for 1 month on the same medium solidified with 0.8% agar. In vitro plantlets were obtained from seeds germinated and cultivated in vitro for 1 month on the callus medium lacking 2,4-dichlorophenoxyacetic acid. Mesophyll pr...

show abstract

Biochemical characterization of thioredoxin 1 from Dictyostelium discoideum

Cited by 20 publications

References 28 publications

The Arabidopsis Plastidial Thioredoxins

The Arabidopsis Plastidial Thioredoxins

In vivo functional discrimination between plant thioredoxins by heterologous expression in the yeast Saccharomyces cerevisiae

Evidence for five divergent thioredoxin h sequences in Arabidopsis thaliana.

Contact Info

Product

Resources

About