Primary Amine-Clustered DNA Aptamer for DNA-Protein Conjugation Catalyzed by Microbial Transglutaminase

Mari Takahara, Rie Wakabayashi, Kosuke Minamihata, Masahiro Goto, Noriho Kamiya

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

DNA-protein conjugates are promising biomolecules for use in areas ranging from therapeutics to analysis because of the dual functionalities of DNA and protein. Conjugation requires site-specific and efficient covalent bond formation without impairing the activity of both biomolecules. Herein, we have focused on the use of a microbial transglutaminase (MTG) that catalyzes the cross-linking reaction between a glutamine residue and a primary amine. In a model bioconjugation, a highly MTG-reactive Gln (Q)-donor peptide (FYPLQMRG, FQ) was fused to enhanced green fluorescent protein (FQ-EGFP) and a primary amine-clustered DNA aptamer was enzymatically synthesized as a novel acyl-acceptor substrate of MTG, whose combination leads to efficient and convenient preparation of DNA-protein conjugates with high purity. Dual functionality of the obtained DNA-EGFP conjugate was evaluated by discrimination of cancer cells via c-Met receptor recognition ability of the DNA aptamer. The DNA aptamer-EGFP conjugate only showed fluorescence toward cells with c-Met overexpression, indicating the retention of the biochemical properties of the DNA and EGFP in the conjugated form.

Original languageEnglish
Pages (from-to)2954-2961
Number of pages8
JournalBioconjugate Chemistry
Volume28
Issue number12
DOIs
Publication statusPublished - Dec 20 2017

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Nucleotide Aptamers
Transglutaminases
Amines
DNA
Proteins
Biomolecules
Covalent bonds
Cross Reactions
Glutamine
Fluorescence
Cells
Peptides
Substrates
Neoplasms

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Cite this

Primary Amine-Clustered DNA Aptamer for DNA-Protein Conjugation Catalyzed by Microbial Transglutaminase. / Takahara, Mari; Wakabayashi, Rie; Minamihata, Kosuke; Goto, Masahiro; Kamiya, Noriho.

In: Bioconjugate Chemistry, Vol. 28, No. 12, 20.12.2017, p. 2954-2961.

Research output: Contribution to journalArticle

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