Chemical synthesis of protein via simultaneous ligation of multiple peptide segments on DNA scaffold

2020 Fiscal Year Final Research Report

• Project/Area Number: 18K05313
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: Nagoya University
• Principal Investigator: Hayashi Gosuke 名古屋大学, 工学研究科, 准教授 (40648268)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: タンパク質化学合成 / ペプチド連結反応 / DNA templated chemistry

Outline of Final Research Achievements

We have developed a novel peptide ligation strategy harnessing the two intrinsic characteristics of oligodeoxynucleotides (ODNs), i.e., their hydrophilicity and hybridization ability, which allowed an increase in the water solubility of peptides and the reaction kinetics due to the proximity effect, respectively. Peptide–ODN conjugates cleavable to regenerate native peptide sequences were synthesized using novel lysine derivatives containing conjugation handles and photolabile linkers, via solid-phase peptide synthesis and subsequent conjugation to 15-mer ODNs. Two complementary conjugates were applied to carbodiimide-mediated peptide ligation on a DNA scaffold and the subsequent DNA removal was conducted by photoirradiation in a traceless fashion.

Free Research Field

生物有機化学

Academic Significance and Societal Importance of the Research Achievements

本研究成果によって、従来mMオーダーで行う必要の合ったペプチド連結反応がサブnMの濃度で実施可能であることを示すことができた。またDNAの親水性によって疎水性ペプチド断片の水溶性向上させ、かつ低濃度で連結反応を行うことで、これまで連結反応に用いることが困難であった疎水性ペプチドを連結することができる新しい技術となった。これにより、これまでより多様なタンパク質を化学的に合成できる可能性があり、ライフサイエンス研究だけでなく、医薬学研究の発展にも寄与することが期待される。