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The consensus motif for N‐myristoylation of plant proteins in a wheat germ cell‐free translation system
Author(s) -
Yamauchi Seiji,
Fusada Naoki,
Hayashi Hidenori,
Utsumi Toshihiko,
Uozumi Nobuyuki,
Endo Yaeta,
Tozawa Yuzuru
Publication year - 2010
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2010.07768.x
Subject(s) - myristoylation , biology , amino acid , biochemistry , translation (biology) , peptide sequence , microbiology and biotechnology , phosphorylation , messenger rna , gene
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.