Structural analysis revealed a novel conformation of the NTRC reductase domain from Chlamydomonas reinhardtii

Giulia Maria Marchetti; Friederike Füsser; Rohit Kumar Singh; Monika Brummel; Oliver Koch; Daniel Kümmel; Michael Hippler

doi:10.1016/j.jsb.2021.107829

Structural analysis revealed a novel conformation of the NTRC reductase domain from Chlamydomonas reinhardtii

J Struct Biol. 2022 Mar;214(1):107829. doi: 10.1016/j.jsb.2021.107829. Epub 2021 Dec 30.

Authors

Giulia Maria Marchetti¹, Friederike Füsser², Rohit Kumar Singh³, Monika Brummel¹, Oliver Koch⁴, Daniel Kümmel⁵, Michael Hippler⁶

Affiliations

¹ Institute of Plant Biology and Biotechnology, University of Münster, 48143 Münster, Germany.
² Institute of Pharmaceutical and Medicinal Chemistry, University of Münster, 48149 Münster, Germany; German Center for Infection Research, University of Münster, 48149 Münster, Germany; Institute of Biochemistry, University of Münster, 48149 Münster, Germany.
³ Institute of Biochemistry, University of Münster, 48149 Münster, Germany; Department of Pharmacology, University of Colorado Denver, USA(1).
⁴ Institute of Pharmaceutical and Medicinal Chemistry, University of Münster, 48149 Münster, Germany; German Center for Infection Research, University of Münster, 48149 Münster, Germany.
⁵ Institute of Biochemistry, University of Münster, 48149 Münster, Germany. Electronic address: daniel.kuemmel@wwu.de.
⁶ Institute of Plant Biology and Biotechnology, University of Münster, 48143 Münster, Germany; Institute of Plant Science and Resources, Okayama University, Kurashiki, Okayama 710-0046, Japan. Electronic address: mhippler@uni-muenster.de.

PMID: 34974142
DOI: 10.1016/j.jsb.2021.107829

Abstract

In plant chloroplasts, thiol regulation is driven by two systems. One relies on the activity of thioredoxins through their light dependent reduction by ferredoxin via a ferredoxin-thioredoxin reductase (FTR). In the other system, a NADPH-dependent redox regulation is driven by a NADPH-thioredoxin reductase C (NTRC). While the thioredoxin system has been deeply studied, a more thorough understanding of the function of this plant specific NTRC is desirable. NTRC is a single polypeptide harbouring a thioredoxin domain (Trx) at the C-terminus of a NADPH-dependent Thioredoxin reductase (TrxR). To provide functional and structural insights, we studied the crystal structure of the TrxR domain of the NTRC from Chlamydomonas reinhardtii (CrNTRC, Cre01.g054150.t1.2) and its Cys136Ser (C136S) mutant, which is characterized by the mutation of the resolving cysteine in the active site of the TrxR domain. Furthermore, we confirmed the role of NTRC as electron donor for 2-Cys peroxiredoxin (PRX) also in C. reinhardtii. The structural data of TrxR were employed to develop a scheme of action which addresses electron transfer between TrxR and Trx of NTRC and between NTRC and its substrates.

Keywords: Chlamydomonas reinhardtii; Crystallography; NTRC; Photosynthesis; Redox; Thioredoxin reductase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis Proteins* / genetics
Arabidopsis* / genetics
Chlamydomonas reinhardtii* / genetics
Chlamydomonas reinhardtii* / metabolism
NADP
Oxidation-Reduction
Oxidoreductases / metabolism
Thioredoxin-Disulfide Reductase / genetics
Thioredoxin-Disulfide Reductase / metabolism
Thioredoxins / metabolism

Substances

Arabidopsis Proteins
Thioredoxins
NADP
Oxidoreductases
NTRC protein, Arabidopsis
Thioredoxin-Disulfide Reductase