광활성 메티오닌 표지단백질 생합성을 위한 메티오닐 tRNA 합성효소 및 이를 이용한 광활성 단백질 G 변이체 제조방법

Abstract

Provided is a methionyl tRNA synthase (MRS) for the biosynthesis of a photomethionine-labeled protein and a method for preparing a photoactivatable protein G variant using same and, more particularly, to an MRS variant in which alanine at the position of 12th is substituted with glycine, leucine at the position of 13th by serine, tyrosine at the position of 260th by phenylalanine, isoleucine at the position of 297th by valine, and histidine at the position of 301st by leucine from the N-terminal of the amino acid sequence of a wild-type Escherichia coli methionyl tRNA synthase. The MRS variant effectively confirms the biosynthesis of a photomethionine (pM)-labeled target protein and thus can be utilized for the biosynthesis of a pM-labeled target protein. In addition, a pM-introduced protein G variant, in which a plasmid encoding the MRS variant (MRS5m) and a PG-C3 plasmid, in which, in the third immunoglobulin G binding region of protein G, positions of 32nd, 35th, and 40th are substituted with a methionine (Met) residue and a position of 37th by an arginine (Arg) residue, are introduced into Escherichia coli and then refined, has a specific covalent bond with an antibody when subject to UV irradiation, and thus the pM-introduced protein G variant using the MRS variant can be utilized for producing a highly sensitive biochip, biosensor, or cell-capturing chip.