Position-specific incorporation of biotinylated non-natural amino acids into a protein in a cell-free translation system.
Biotinylation is useful for the detection, purification and immobilization of proteins. It is performed by chemical modification, although position-specific and quantitative biotinylation is rarely achieved. We developed a position-specific biotinylation method using biotinylated non-natural amino acids. We showed that biotinylated p-aminophenylalanine derivatives were incorporated into a protein more efficiently than biotinylated lysine derivatives in a cell-free translation system. In addition, the biotinylated p-aminophenylalanines overcame the serious position-dependency observed for biotinylated lysines. The present method will be useful for detection and purification of proteins along with comprehensive exploration of surface-exposed residues and oriented immobilization of proteins.
Watanabe
Takayoshi
T
School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa 923-1292, Japan.
Muranaka
Norihito
N
Iijima
Issei
I
Hohsaka
Takahiro
T
D001665:Binding Sites; D001710:Biotin / Q000737:chemistry*; D019921:Biotinylation / Q000379:methods; D002474:Cell-Free System / Q000378:metabolism; D003062:Codon; D008239:Lysine / Q000737:chemistry; D008958:Models, Molecular; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000737:chemistry; D014176:Protein Biosynthesis; D019809:Streptavidin / Q000378:metabolism
D003062:Codon; C009722:4-aminophenylalanine; D010649:Phenylalanine; D001710:Biotin; D019809:Streptavidin; D008239:Lysine