TY - JOUR
T1 - Transglutaminase-mediated internal protein labeling with a designed peptide loop
AU - Mori, Yutaro
AU - Goto, Masahiro
AU - Kamiya, Noriho
N1 - Funding Information:
This research was supported by the Research for Promoting Technological Seeds from the Japan Science and Technology Agency (JST) of Japan and partly by a Grant-in-Aid for the Global COE Program, ‘Science for Future Molecular Systems’ from the MEXT of Japan.
PY - 2011/7/15
Y1 - 2011/7/15
N2 - Post-translational internal protein labeling was explored through the insertion of a 13-mer peptidyl loop specifically recognized by microbial transglutaminase (MTG). The peptidyl loop included one lysine residue (abbreviated as the K-loop), and was designed and inserted into two different regions of the protein bacterial alkaline phosphatase (BAP). MTG-mediated selective labeling of a lysine residue in the K-loop was achieved with a functional Gln-donor substrate. Internal protein labeling in the vicinity of the active site of BAP (residues 91-93) markedly decreased the activity of the enzyme. Conversely, insertion of the K-loop at a site distal from the active site (residues 219-221) afforded site-specific and covalent internal protein labeling without impairing the activity of the enzyme.
AB - Post-translational internal protein labeling was explored through the insertion of a 13-mer peptidyl loop specifically recognized by microbial transglutaminase (MTG). The peptidyl loop included one lysine residue (abbreviated as the K-loop), and was designed and inserted into two different regions of the protein bacterial alkaline phosphatase (BAP). MTG-mediated selective labeling of a lysine residue in the K-loop was achieved with a functional Gln-donor substrate. Internal protein labeling in the vicinity of the active site of BAP (residues 91-93) markedly decreased the activity of the enzyme. Conversely, insertion of the K-loop at a site distal from the active site (residues 219-221) afforded site-specific and covalent internal protein labeling without impairing the activity of the enzyme.
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U2 - 10.1016/j.bbrc.2011.06.073
DO - 10.1016/j.bbrc.2011.06.073
M3 - Article
C2 - 21703236
AN - SCOPUS:79960288573
VL - 410
SP - 829
EP - 833
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 4
ER -