Establishment of the in vitro docking (IVD) method for the functional expression of human membrane proteins on magnetic particles

2016 Fiscal Year Final Research Report

• Project/Area Number: 26289308
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: Yoshino Tomoko 東京農工大学, 工学(系)研究科(研究院), 准教授 (30409750)
• Co-Investigator(Kenkyū-buntansha): 前田 義昌 東京農工大学, 工学(系)研究科(研究院), 助教 (30711155)
• Co-Investigator(Renkei-kenkyūsha): TANAKA TSUYOSHI 東京農工大学, 大学院工学研究院, 教授 (20345333)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 磁性粒子 / 膜タンパク質 / in vitro docking / 細胞内区画

Outline of Final Research Achievements

This study aimed to establish the in vitro docking (IVD) method in the magnetotactic bacterium towards the development of the functional composites of bacterial magnetic particles (BacMPs) and human-derived membrane proteins. In the IVD method, recombinant proteins (including membrane proteins) expressed in the periplasmic space of the magnetotactic bacterium were assembled onto BacMPs, which were separately synthesized in the identical bacterium, with the aid of docking protein pairs after cell disruption. Several types of docking protein pairs were examined for the IVD method. Subsequently, as a proof-of-concept, thyroid stimulating hormone receptor (TSHR) was displayed on the BacMPs through the IVD method. In addition, the minimum expression system was established in the magnetotactic bacterium for appropriate protein folding, which could be beneficial for functional expression of recombinant proteins.

Free Research Field

生物機能・バイオプロセス