Abstract
Transglutaminase (TGase) is an enzyme that catalyzes the post-translational covalent cross-linking of Gln- and Lys-containing peptides and/or proteins according to its substrate specificity. We have recently designed a variety of Gln-donor fluorescent substrates of microbial transglutaminase (MTG) from Streptomyces mobaraensis and evaluated their potential use in MTG-mediated covalent protein labeling. The newly designed substrates are based on the relatively broad substrate recognition of MTG for the substitution of the N-terminal group of a conventional TGase substrate, benzyloxycarbonyl-l-glutaminylglycine (Z-QG). It is revealed that MTG is capable of accepting a diverse range of fluorophores in place of the N-terminal moiety of Z-QG when linked via a suitable linker. Here, we show the potential utility of a new fluorescent substrate for peptide tag-directed covalent protein labeling by employing fluorescein-4-isothiocyanate-β-Ala-QG as a model Gln-donor substrate for MTG.
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Acknowledgments
We are grateful to Ajinomoto Co., Inc. (Japan) for providing us with the MTG sample. This work was supported by the Research for Promoting Technological Seeds from the Japan Science and Technology Agency (JST) of Japan, and also in part by a Grant-in-Aid for the Global COE Program, “Science for Future Molecular Systems” from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan.
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Kamiya, N., Abe, H. (2011). New Fluorescent Substrates of Microbial Transglutaminase and Its Application to Peptide Tag-Directed Covalent Protein Labeling. In: Mark, S. (eds) Bioconjugation Protocols. Methods in Molecular Biology, vol 751. Humana Press. https://doi.org/10.1007/978-1-61779-151-2_7
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DOI: https://doi.org/10.1007/978-1-61779-151-2_7
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