Method for enhancement of plant redox-related protein expression and its application for in vitro reduction of chloroplastic thioredoxins

Ken Motohashi, Yuki Okegawa

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

Plant redox-related proteins were overexpressed using a genetic codon substitution downstream of the translation initiation codon. This method significantly improved recombinant protein expression levels of Arabidopsis chloroplastic thioredoxins and cytosolic nicotinamide adenine dinucleotide phosphate (NADPH)-dependent thioredoxin reductase (E.C. 1.8.1.9) in Escherichia coli. Using these proteins, the in vitro chloroplastic thioredoxins-reduction system was reconstituted in an NADPH-dependent manner. This system could convert the five classes of chloroplastic Arabidopsis thioredoxins and two chloroplastic Spinach thioredoxins to their reduced forms, independent of dithiothreitol and the photosynthetic electron transport system.

Original languageEnglish
Pages (from-to)152-156
Number of pages5
JournalProtein Expression and Purification
Volume101
DOIs
Publication statusPublished - Sep 2014
Externally publishedYes

Keywords

  • Chloroplast
  • Disulfide bond
  • NADPH-dependent thioredoxin reductase
  • Redox regulation
  • Thioredoxin
  • Translation initiation codon

ASJC Scopus subject areas

  • Biotechnology

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