TY - JOUR
T1 - Primary Amine-Clustered DNA Aptamer for DNA-Protein Conjugation Catalyzed by Microbial Transglutaminase
AU - Takahara, Mari
AU - Wakabayashi, Rie
AU - Minamihata, Kosuke
AU - Goto, Masahiro
AU - Kamiya, Noriho
N1 - Funding Information:
This work was financially supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI (No. JP16H04581 to N. K.). M.T. was supported by a Research Fellowship from the JSPS for Young Scientists with a Grant-in-Aid for JSPS Fellows (No. 26-4260) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan. Part of this research was supported by the Nanotechnology Platform Project (Molecules and Materials Synthesis) from MEXT of Japan. We thank the Edanz Group (www.edanzediting.com/ac) for editing a draft of this manuscript.
Publisher Copyright:
© 2017 American Chemical Society.
PY - 2017/12/20
Y1 - 2017/12/20
N2 - 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.
AB - 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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U2 - 10.1021/acs.bioconjchem.7b00594
DO - 10.1021/acs.bioconjchem.7b00594
M3 - Article
C2 - 29131594
AN - SCOPUS:85038573332
VL - 28
SP - 2954
EP - 2961
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
SN - 1043-1802
IS - 12
ER -