Cell-free synthesis of zinc-binding proteins

Takayoshi Matsuda, Takanori Kigawa, Seizo Koshiba, Makoto Inoue, Masaaki Aoki, Kazuhiko Yamasaki, Motoaki Seki, Kazuo Shinozaki, Shigeyuki Yokoyama

Research output: Contribution to journalArticlepeer-review

30 Citations (Scopus)

Abstract

Cell-free protein synthesis has become one of the standard methods for protein expression. The cell-free method is suitable for the synthesis of a protein that requires a ligand for its enzymatic activity and/or structure formation and stabilization, since it is an open system, which allows us to add the proper ligand to the reaction mixture. A large number of proteins that require zinc for their function are involved in diverse cellular processes, including transcription, DNA replication, metabolism, and cell signaling. In this study, we analyzed the effects of zinc on the cell-free synthesis of plant-specific zinc-binding transcription factors. The solubility and/or stability of the proteins were significantly increased in the presence of the proper concentration of zinc during the cell-free reaction. NMR analyses confirmed that correctly folded proteins were synthesized by the cell-free method. These results indicate that the cell-free method can be used to synthesize correctly folded and functional zinc-binding proteins.

Original languageEnglish
Pages (from-to)93-100
Number of pages8
JournalJournal of Structural and Functional Genomics
Volume7
Issue number2
DOIs
Publication statusPublished - 2006 Jun
Externally publishedYes

Keywords

  • Arabido psis thaliana
  • Cell-free protein synthesis
  • In vitro translation
  • Structural genomics
  • Zinc-binding protein

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Genetics

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