Fluorescent substrates for covalent protein labeling catalyzed by microbial transglutaminase

Noriho Kamiya, Hiroki Abe, Masahiro Goto, Yukiko Tsuji, Hiroyuki Jikuya

Research output: Contribution to journalArticle

17 Citations (Scopus)

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 languageEnglish
Pages (from-to)3407-3412
Number of pages6
JournalOrganic and Biomolecular Chemistry
Volume7
Issue number17
DOIs
Publication statusPublished - Sep 25 2009

Fingerprint

Transglutaminases
Labeling
marking
proteins
Substrates
Fluorophores
Proteins
glutaminyl-glycine
phosphatases
Aminocaproic Acid
alanine
rhodamine
Rhodamines
peptides
flexibility
Fluorescein
Alanine
Alkaline Phosphatase
substitutes
Substitution reactions

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 journalArticle

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