Suppression of oxidative stress by protein-protein interaction between cytochrome P450 and UDP-glucuronosyltransferase

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K16045
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 47060:Clinical pharmacy-related
• Research Institution: Sojo University
• Principal Investigator: Miyauchi Yuu 崇城大学, 薬学部, 講師 (50799947)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: シトクロムP450 / UDP-グルクロン酸転移酵素 / タンパク質間相互作用 / バキュロウイルス / 哺乳動物細胞 / 活性酸素種 / 酸化ストレス / 糖化産物

Outline of Final Research Achievements

We have reported that a protein-protein interaction between different kinds of drug-metabolizing enzymes, cytochrome P450 (CYP) and UDP-glucuronosyltransferase (UGT) work as one of the post-translational regulators of their enzymatic functions. For further analysis of the CYP-UGT interaction, we hypothesized that the CYP-UGT interaction suppresses the CYP-mediated production of reactive oxygen species, which is one of the physiological roles of the interaction. To analyze the CYP-UGT interaction in mammalian cells, we have constructed several expression systems in this study.
A baculovirus-mammalian cell expression system (bac-mam system) is applied to the expression of drug-metabolizing enzyme for the first time, and expressions of the targeted CYP and UGT isoforms were confirmed in COS-1 cells, which was published in an international journal.
We hope that the expression systems constructed in this study accelerate our study of CYP-UGT interaction in the future.

Free Research Field

衛生薬学、薬物代謝、毒性学

Academic Significance and Societal Importance of the Research Achievements

本研究では生理的な条件に近づけてCYP-UGT相互作用を解析するため、哺乳動物細胞にCYPとUGTを発現させた。一般的な発現プラスミドとトランスフェクション試薬を用いる方法では、CYPとUGTの発現量の制御が難しい。本研究で新たに用いたbac-mam systemでは、遺伝子改変バキュロウイルスを用いて哺乳動物細胞に遺伝子導入を行う。バキュロウイルスは宿主が昆虫細胞に限定されるため他のウイルスと比べて安全性が高く、また遺伝子導入が困難な肝細胞への応用も見込まれる。本研究成果はCYP-UGT相互作用の研究に限らず、発現系の選択肢を広げることで他の研究にも波及効果が期待される。