Production of peptide that associate to the dimer interface of kinase domains

2020 Fiscal Year Final Research Report

• Project/Area Number: 18K05359
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 37030:Chemical biology-related
• Research Institution: Kumamoto University
• Principal Investigator: Kobashigawa Yoshihiro 熊本大学, 大学院生命科学研究部(薬), 准教授 (90455600)
• Co-Investigator(Kenkyū-buntansha): 森岡 弘志 熊本大学, 大学院生命科学研究部(薬), 教授 (20230097)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 線維芽細胞増殖因子受容体 / チロシンキナーゼ / 二量体

Outline of Final Research Achievements

Receptor tyrosine kinases transiently form inter-kinase domain dimers during activation. In this study, we attempted to search for peptides that bind to the dimer interface between kinase domains of fibroblast growth factor receptor 1 (FGFR1). The helix G plays an important role in the interaction between kinase domains of FGFR1. Therefore, an interaction residue between kinase domains of helix G was introduced into one of the helices of a peptide that forms helix-loop-helix structure (HLH peptide). As a result, it was found that this peptide binds to the kinase domain of FGFR1 and enhances the auto-phosphorylation of FGFR1. The HLH peptide was prepared as a fusion protein with maltose-binding protein (MBP). It was also revealed that inexpensive corn starch can be used as an affinity chromatography carrier for the purification of the MBP fusion proteins.

Free Research Field

構造生物化学

Academic Significance and Societal Importance of the Research Achievements

本研究では、受容体型チロシンキナーゼを活性化するペプチドを見出した。ペプチドの生産には大腸菌発現系を用いたが、ペプチド鎖長上は化学合成でも生産可能である。受容体型チロシンキナーゼを活性化するタンパク質 (成長因子・細胞増殖因子)は細胞培養の際に試薬として使用される。それらは、複雑な構造を有し、生産ロット間の品質のばらつきにつながる。今後、さらなる研究は必要だが、受容体型チロシンキナーゼを活性化するペプチドを化学合成したものが代替できれば、生産ロット間の品質格差の抑制につながる。