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

Authors
  • Takahara, Mari1
  • Wakabayashi, Rie1
  • Minamihata, Kosuke1
  • Goto, Masahiro1
  • Kamiya, Noriho1
  • 1 Department of Applied Chemistry, Graduate School of Engineering and ‡Division of Biotechnology, Center for Future Chemistry, Kyushu University , 744 Motooka, Fukuoka 819-0395, Japan. , (Japan)
Type
Published Article
Journal
Bioconjugate Chemistry
Publisher
American Chemical Society
Publication Date
Dec 20, 2017
Volume
28
Issue
12
Pages
2954–2961
Identifiers
DOI: 10.1021/acs.bioconjchem.7b00594
PMID: 29131594
Source
Medline
License
Unknown

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.

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