Comprehensive screening of drug-binding proteins using a detection method for their interactions based on chemical reactions between reactive peptide tags

2021 Fiscal Year Final Research Report

• Project/Area Number: 18K05348
• 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: Gunma University
• Principal Investigator: Takahashi Tsuyoshi 群馬大学, 大学院理工学府, 准教授 (90345380)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Keywords: オフターゲットタンパク質 / ドラッグ・リポジショニング / βラクタマーゼ

Outline of Final Research Achievements

In the present study, IDNCL-ER, a detection system for ligand-protein interactions, has been improved to discover the proteins that bind to drugs and drug candidates. To this purpose, we employed β-lactamase, which is known as an enzyme to break β-lactam ring, as a reporter enzyme. A split intein, which can self-catalyze protein trans-splicing (PTS) reaction, was also employed to develop the detection method. In the detection method, a synthetic ligand such as atorvastatin, a known cholesterol-lowering drug, was conjugated with the short peptide tag containing the C-terminal sequences of split intein and β-lactamase. When the synthetic ligand binds to a target protein, PDE6D, bearing N-terminal sequences β-lactamase and split intein, PTS reaction occurs and active β-lactamase is generated. Using this method, we have assessed the interactions between atorvastatin and PDE6D in vitro, and in E. coli.

Free Research Field

ペプチド工学

Academic Significance and Societal Importance of the Research Achievements

本研究は、薬剤などの化合物に結合するタンパク質を網羅的に探索する方法を開発する基礎的研究である。開発した方法を用いて、特定の薬剤に結合するオフターゲットタンパク質の探索が可能となれば、副作用などの原因の解明に繋がる可能性が期待できる。それにより、副作用を低減する薬剤開発なども進展すると期待できる。このように本研究は、学術的かつ社会的に意義のある研究であると考えられる。