Structure of Hordeum vulgare NADPH-dependent thioredoxin reductase 2. Unwinding the reaction mechanism

Kristine Groth Kirkensgaard, Per Hägglund, Christine Finnie, Birte Svensson, A. Henriksen

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    Abstract

    Thioredoxins (Trxs) are protein disulfide reductases that regulate the intracellular redox environment and are important for seed germination in plants. Trxs are in turn regulated by NADPH-dependent thioredoxin reductases (NTRs), which provide reducing equivalents to Trx using NADPH to recycle Trxs to the active form. Here, the first crystal structure of a cereal NTR, HvNTR2 from Hordeum vulgare (barley), is presented, which is also the first structure of a monocot plant NTR. The structure was determined at 2.6 A resolution and refined to an R (cryst) of 19.0% and an R (free) of 23.8%. The dimeric protein is structurally similar to the structures of AtNTR-B from Arabidopsis thaliana and other known low-molecular-weight NTRs. However, the relative position of the two NTR cofactor-binding domains, the FAD and the NADPH domains, is not the same. The NADPH domain is rotated by 25 degrees and bent by a 38% closure relative to the FAD domain in comparison with AtNTR-B. The structure may represent an intermediate between the two conformations described previously: the flavin-oxidizing (FO) and the flavin-reducing (FR) conformations. Here, analysis of interdomain contacts as well as phylogenetic studies lead to the proposal of a new reaction scheme in which NTR-Trx interactions mediate the FO to FR transformation.
    Original languageEnglish
    JournalActa Crystallographica. Section D: Biological Crystallography
    Volume65
    Pages (from-to)932-941
    ISSN0907-4449
    DOIs
    Publication statusPublished - 2009

    Keywords

    • redox regulation
    • NADPH
    • barley
    • seed development
    • disulfide oxidoreductases
    • NADPH-dependent thioredoxin reductases
    • germination

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