Abstract
Novel small substrates with a variety of fluorophores were designed for the covalent labeling of proteins catalyzed by microbial transglutaminase (MTG). The new design is based on the flexibility in the substrate recognition of MTG for the substitution of the N-terminal protecting group of a conventional transglutaminase substrate, benzyloxycarbonyl-L-glutaminylglycine (Z-QG). Here we report for the first time that MTG can accept diverse fluorophores (dansyl, fluorescein, and rhodamine derivatives) in place of the benzyloxycarbonyl moiety when linked via a β-alanine or ε-aminocaproic acid linker. The utility of the new fluorescent substrates was demonstrated by site-specific, covalent and quantitative labeling of an MTG-reactive Lys-containing peptide tag fused to the N-terminus of a recombinant bacterial alkaline phosphatase with retention of target protein functionality.
| Original language | English |
|---|---|
| Pages (from-to) | 3407-3412 |
| Number of pages | 6 |
| Journal | Organic and Biomolecular Chemistry |
| Volume | 7 |
| Issue number | 17 |
| DOIs | |
| Publication status | Published - Sep 25 2009 |
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All Science Journal Classification (ASJC) codes
- Physical and Theoretical Chemistry
- Organic Chemistry
- Biochemistry
Cite this
Fluorescent substrates for covalent protein labeling catalyzed by microbial transglutaminase. / Kamiya, Noriho; Abe, Hiroki; Goto, Masahiro; Tsuji, Yukiko; Jikuya, Hiroyuki.
In: Organic and Biomolecular Chemistry, Vol. 7, No. 17, 25.09.2009, p. 3407-3412.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Fluorescent substrates for covalent protein labeling catalyzed by microbial transglutaminase
AU - Kamiya, Noriho
AU - Abe, Hiroki
AU - Goto, Masahiro
AU - Tsuji, Yukiko
AU - Jikuya, Hiroyuki
PY - 2009/9/25
Y1 - 2009/9/25
N2 - Novel small substrates with a variety of fluorophores were designed for the covalent labeling of proteins catalyzed by microbial transglutaminase (MTG). The new design is based on the flexibility in the substrate recognition of MTG for the substitution of the N-terminal protecting group of a conventional transglutaminase substrate, benzyloxycarbonyl-L-glutaminylglycine (Z-QG). Here we report for the first time that MTG can accept diverse fluorophores (dansyl, fluorescein, and rhodamine derivatives) in place of the benzyloxycarbonyl moiety when linked via a β-alanine or ε-aminocaproic acid linker. The utility of the new fluorescent substrates was demonstrated by site-specific, covalent and quantitative labeling of an MTG-reactive Lys-containing peptide tag fused to the N-terminus of a recombinant bacterial alkaline phosphatase with retention of target protein functionality.
AB - Novel small substrates with a variety of fluorophores were designed for the covalent labeling of proteins catalyzed by microbial transglutaminase (MTG). The new design is based on the flexibility in the substrate recognition of MTG for the substitution of the N-terminal protecting group of a conventional transglutaminase substrate, benzyloxycarbonyl-L-glutaminylglycine (Z-QG). Here we report for the first time that MTG can accept diverse fluorophores (dansyl, fluorescein, and rhodamine derivatives) in place of the benzyloxycarbonyl moiety when linked via a β-alanine or ε-aminocaproic acid linker. The utility of the new fluorescent substrates was demonstrated by site-specific, covalent and quantitative labeling of an MTG-reactive Lys-containing peptide tag fused to the N-terminus of a recombinant bacterial alkaline phosphatase with retention of target protein functionality.
UR - http://www.scopus.com/inward/record.url?scp=70349283056&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=70349283056&partnerID=8YFLogxK
U2 - 10.1039/b904046c
DO - 10.1039/b904046c
M3 - Article
C2 - 19675894
AN - SCOPUS:70349283056
VL - 7
SP - 3407
EP - 3412
JO - Organic and Biomolecular Chemistry
JF - Organic and Biomolecular Chemistry
SN - 1477-0520
IS - 17
ER -