tailieunhanh - Báo cáo khoa học: The consensus motif for N-myristoylation of plant proteins in a wheat germ cell-free translation system

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 ofArabidopsis thalianaproteins. | The consensus motif for N-myristoylation of plant proteins in a wheat germ cell-free translation system Seiji Yamauchi1 Naoki Fusada1 2 Hidenori Hayashi1 Toshihiko Utsumi3 Nobuyuki Uozumi4 Yaeta Endo1 5 and Yuzuru Tozawa1 1 Cell-Free Science and Technology Research Center and Venture Business Laboratory Ehime University Matsuyama Japan 2 Department of Applied BiologicalScience College of Bioresource Sciences Nihon University Fujisawa Japan 3 Department of BiologicalChemistry Faculty of Agriculture Yamaguchi University Japan 4 Department of Biomolecular Engineering Graduate Schoolof Engineering Tohoku University Sendai Japan 5 Systems and StructuralBiology Center RIKEN Yokohama Japan Keywords cell-free translation myristoylation N-myristoyltransferase plant wheat germ Correspondence Y. Tozawa Division of Biomolecular Engineering Cell-Free Science and Technology Research Center Ehime University 3 Bunkyo-cho Matsuyama Ehime 790-8577 Japan Fax 81 89 927 8528 Tel 81 89 927 8274 E-mail tozaway@ Received 10 May 2010 revised 4 July 2010 accepted 8 July 2010 doi 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-myristoy-lation 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