Abstract
Lipid-modified proteins play critical roles in regulating many cellular functions. Myristoylation is well-recognized form of lipid modification in eukaryote. In this process, myristate is attached to the N-terminus of the proteins. In the last conference, we have reported the construction of myristoylation system in wheat-embryo cell-free translation system. To determine the sequence specificity in the myristoylation consensus motif for plant, we next analyzed the combination of amino acids at position 3 and 6. We used G protein γ-subunit from Arabidopsis thaliana (AGG1) fused the myristoylation consensus motifs, MGXAA(A/S)AAAA (X shows each 20 amino acid.), as substrates and the susceptibility of these proteins to myristoylation was estimated by using wheat-embryo cell-free translation system. When Ser was located at position 6, 12 amino acids were permitted at position 3 to direct myristoylation, while only 2 amino acids (Asn and Gln) were permitted at position 3 when Ala was located at position 6. To obtain further information, analyses of the amino acids at other positions and removal of the initiating Met are now in progress.
