The consensus motif for N-myristoylation of plant proteins in a wheat germ cell-free translation system

Seiji Yamauchi, Naoki Fusada, Hidenori Hayashi, Toshihiko Utsumi, Nobuyuki Uozumi, Yaeta Endo, Yuzuru Tozawa

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Protein N-myristoylation plays key roles in various cellular functions in eukaryotic organisms. To clarify the relationship between the efficiency of protein N-myristoylation and the amino acid sequence of the substrate in plants, we have applied a wheat germ cell-free translation system with high protein productivity to examine the N-myristoylation of various wild-type and mutant forms of Arabidopsis thaliana proteins. Evaluation of the relationship between removal of the initiating Met and subsequent N-myristoylation revealed that constructs containing Pro at position 3 do not undergo N-myristoylation, primarily because of an inhibitory effect of this amino acid on elimination of the initiating Met by methionyl aminopeptidase. Our analysis of the consensus sequence for N-myristoylation in plants focused on the variability of amino acids at positions 3, 6 and 7 of the motif. We found that not only Ser at position 6 but also Lys at position 7 affects the selectivity for the amino acid at position 3. The results of our analyses allowed us to identify several A. thaliana proteins as substrates for N-myristoylation that had previously been predicted not to be candidates for such modification with a prediction program. We have thus shown that a wheat germ cell-free system is a useful tool for plant N-myristoylome analysis. This in vitro approach will facilitate comprehensive determination of N-myristoylated proteins in plants.

Original languageEnglish
Pages (from-to)3596-3607
Number of pages12
JournalFEBS Journal
Volume277
Issue number17
DOIs
Publication statusPublished - 2010 Sep

Keywords

  • N-myristoyltransferase
  • cell-free translation
  • myristoylation
  • plant
  • wheat germ

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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