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N‐terminal glycine‐specific protein conjugation catalyzed by microbial transglutaminase
Author(s) -
Tanaka Tsutomu,
Kamiya Noriho,
Nagamune Teruyuki
Publication year - 2005
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2005.02.064
Subject(s) - biochemistry , dihydrofolate reductase , chemistry , residue (chemistry) , peptide , n terminus , green fluorescent protein , epitope , amino acid , tissue transglutaminase , oligopeptide , peptide sequence , glycine , mutagenesis , enzyme , biology , mutant , gene , antigen , genetics
Here, we report the N‐terminal glycine (Gly) residue of a target protein can be a candidate primary amine for site‐specific protein conjugation catalyzed by microbial transglutaminase (MTG) from Streptomyces mobaraensis . Gly5‐enhanced green fluorescent protein (EGFP) (EGFP with five additional Gly residues at its N‐terminus) was cross‐linked with Myc‐dihydrofolate reductase (DHFR) (DHFR with the myc epitope sequence at its N‐terminus) to yield DHFR–EGFP heterodimers. The reactivities of additional peptidyl linkers were investigated and the results obtained suggested that at least three additional Gly residues at the N‐terminus were required to yield the EGFP–DHFR heterodimeric form. Site‐directed mutagenesis analysis revealed marked preference of MTG for amino acids adjacent to the N‐terminal Gly residue involved in the protein conjugation. In addition, peptide–protein conjugation was demonstrated by MTG‐catalyzed N‐terminal Gly‐specific modification of a target protein with the myc epitope peptide.