De-novo protein production technologies for evaluation and regulation of protein lifetime
Associate professor, Nagoya University Department of Biomolecular Engineering, Graduate School of Engineering
https://www.chembio.nagoya-u.ac.jp/labhp/bioanal2/en/
researchmap: https://researchmap.jp/ghayashi
We aim to establish methodologies that can evaluate and regulate protein lifetime by utilizing state-of-the-art de-novo protein production techniques based on “chemical protein synthesis” and “in vitro selection”. Chemical protein synthesis enables us to synthesize proteins with various post-translational modifications including mono-ubiquitin and Ub chains, which are used in biochemical and structural analyses. On the other hand, “in vitro selection” can produce antibody-like proteins (ALPs) such as nanobody and monobody that selectively bind to proteins of interest. These ALPs could be applied to a new bio-degrader by dimerizing two different APLs. Furthermore, we will combine these two techniques to obtain ALPs that bind to chemically synthesized ubiquitinated proteins. The chemically synthesized proteins and/or ALPs produced in this study will be provided to collaborators for biochemical analysis, structural analysis, or live-cell analysis.
- Nakatsu K., Okamoto A., Hayashi G., & Murakami H. (2022) Repetitive Thiazolidine Deprotection Using a Thioester‐Compatible Aldehyde Scavenger for One‐Pot Multiple Peptide Ligation**. Angewandte Chemie International Edition. 61, e202206240
- Kamo N., Kujirai T., Kurumizaka H., Murakami H., Hayashi G., & Okamoto A. (2021) Organoruthenium-catalyzed chemical protein synthesis to elucidate the functions of epigenetic modifications on heterochromatin factors. Chem Sci. 12, 5926–5937
- Kondo T., Iwatani Y., Matsuoka K., Fujino T., Umemoto S., Yokomaku Y., Ishizaki K., Kito S., Sezaki T., Hayashi G., & Murakami H. (2020) Antibody-like proteins that capture and neutralize SARS-CoV-2. Sci Adv. 10.1126/sciadv.abd3916
- Yanase M., Nakatsu K., Cardos C. J., Konda Y., Hayashi G., & Okamoto A. (2019) Cysteinylprolyl imide (CPI) peptide: a highly reactive and easily accessible crypto-thioester for chemical protein synthesis. Chem Sci. 10, 5967–5975
- Kamo N., Hayashi G., & Okamoto A. (2018) Triple Function of 4‐Mercaptophenylacetic Acid Promotes One‐Pot Multiple Peptide Ligation. Angewandte Chemie. 130, 16771–16775