Insights into the function of NADPH thioredoxin reductase C (NTRC) based on identification of NTRC-interacting proteins in vivo.
Chloroplast
NTRC
TAP-Tag
peroxiredoxin
proteomics
redox regulation
Journal
Journal of experimental botany
ISSN: 1460-2431
Titre abrégé: J Exp Bot
Pays: England
ID NLM: 9882906
Informations de publication
Date de publication:
24 10 2019
24 10 2019
Historique:
received:
08
03
2019
accepted:
04
07
2019
pubmed:
12
7
2019
medline:
18
8
2020
entrez:
12
7
2019
Statut:
ppublish
Résumé
Redox regulation in heterotrophic organisms relies on NADPH, thioredoxins (TRXs), and an NADPH-dependent TRX reductase (NTR). In contrast, chloroplasts harbor two redox systems, one that uses photoreduced ferredoxin (Fd), an Fd-dependent TRX reductase (FTR), and TRXs, which links redox regulation to light, and NTRC, which allows the use of NADPH for redox regulation. It has been shown that NTRC-dependent regulation of 2-Cys peroxiredoxin (PRX) is critical for optimal function of the photosynthetic apparatus. Thus, the objective of the present study was the analysis of the interaction of NTRC and 2-Cys PRX in vivo and the identification of proteins interacting with them with the aim of identifying chloroplast processes regulated by this redox system. To assess this objective, we generated Arabidopsis thaliana plants expressing either an NTRC-tandem affinity purification (TAP)-Tag or a green fluorescent protein (GFP)-TAP-Tag, which served as a negative control. The presence of 2-Cys PRX and NTRC in complexes isolated from NTRC-TAP-Tag-expressing plants confirmed the interaction of these proteins in vivo. The identification of proteins co-purified in these complexes by MS revealed the relevance of the NTRC-2-Cys PRX system in the redox regulation of multiple chloroplast processes. The interaction of NTRC with selected targets was confirmed in vivo by bimolecular fluorescence complementation (BiFC) assays.
Identifiants
pubmed: 31294455
pii: 5531050
doi: 10.1093/jxb/erz326
pmc: PMC6812714
doi:
Substances chimiques
Arabidopsis Proteins
0
Green Fluorescent Proteins
147336-22-9
NTRC protein, Arabidopsis
EC 1.8.1.9
Thioredoxin-Disulfide Reductase
EC 1.8.1.9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
5787-5798Informations de copyright
© The Author(s) 2019. Published by Oxford University Press on behalf of the Society for Experimental Biology.
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