Characterization of thioredoxin-like 3, a conserved protein in the green lineage
2024
de Bont, Linda | Dhalleine, Tiphaine | Torresi, Florencia | Donnay, Natacha | Mathiot, Sandrine | Didierjean, Claude | Rouhier, Nicolas | Interactions Arbres-Microorganismes (IAM) ; Université de Lorraine (UL)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Universidad Nacional de Rosario [Santa Fe] | Cristallographie, Résonance Magnétique et Modélisations (CRM2) ; Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS) | LabEx Arbre | Pierre Frendo
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Afficher plus [+] Moins [-]anglais. Thioredoxins (TRX) are ubiquitous proteins regulating the redox state of numerous proteins, in particular in chloroplasts. These small proteins have a particular fold and a CxxC motif comprising two active-site cysteines. A predicted chloroplastic protein possessing these characteristics has been automatically annotated as TRX-like but its function is unknown and its predicted fold is more related to thiol peroxidases (i.e. AhpC_TSA2-like domain). It is present in microalgae and land plants including trees and we referred it to as TRX-like3. The recombinant TRX-like3 proteins from Arabidopsis thaliana and Chlamydomonas reinhardtii have been expressed in Escherichia coli and purified. The structure of AtTRX-like3 has been solved. Like regular TRXs, it is a monomer and the CxxC motif present in the first -helix is surface exposed. However, AtTRX-like3 possesses an elongated N-terminal region and an insertion between the β3 and β4 strands compared to conventional TRXs. While both TRX-like3 are oxidized by H2O2, no thiol peroxidase activity has been detected. A reductase activity was not detected either, using insulin as a substrate. Since the biochemical analyses do not give information about a possible function as a reductase, Chlamydomonas mutants have been selected. Preliminary results indicate that Trx-like3 mutants exhibit a growth retardation on a minimal medium under light which was not visible when grown photo-autotrophically on rich-media. This opens the way for a deeper functional analysis.
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