title: A novel immobilization method of an active protein via in vitro N-terminal specific incorporation system of nonnatural amino acids.
abstract: Recently, we succeeded in incorporating a biotin tag into an active protein, but only at the N-terminal site, in the presence of an Escherichia coli initiator tRNA(fMet) aminoacylated with methionine biotinylated at the alpha-amino group. The biotinylated protein was immobilized on a streptavidin-matrix. We have tried to increase the biotin labeling efficiency (immobilization efficiency) by decreasing concentrations of non-biotinylated tRNA(fMet)s in the translation system.
authors.affiliation: National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba Science City, 305-8562, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Taki
authors.forename: S Y
authors.initials: SY
authors.lastname: Sawata
authors.forename: K
authors.initials: K
authors.lastname: Taira
mesh_terms: D019921:Biotinylation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D008715:Methionine / Q000737:chemistry*; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry; D012358:RNA, Transfer, Met / Q000138:chemical synthesis / Q000737:chemistry*; D019809:Streptavidin / Q000737:chemistry
chemical_list: D011506:Proteins; D012358:RNA, Transfer, Met; D019809:Streptavidin; D008715:Methionine