Structure analysis of monooxygenase using XFEL and microfluidic device

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K22403
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Sugimoto Hiroshi 国立研究開発法人理化学研究所, 放射光科学研究センター, 専任研究員 (90344043)
• Co-Investigator(Kenkyū-buntansha): 真栄城 正寿 北海道大学, 工学研究院, 助教 (40744248)
• Project Period (FY): 2019-06-28 – 2021-03-31
• Keywords: ヘム / X線結晶構造解析 / X線自由電子レーザー

Outline of Final Research Achievements

Monooxygenase reactions catalyzed by cytochrome P450 family require the sequential input of two reducing equivalents (i.e., two electrons and two protons) which activate the oxygen molecule. We developed a prototype of device to control such a multi-step reaction of monooxygenase in crystalline phase. We performed a feasibility study of X-ray diffraction experiments using this device with microcrystals of cytochrome P450 at X-ray free electron laser (XFEL) facility. The result has shown that the device is useful for data collection system of serial crystallography using XFEL and photo-caged reagents, which pave the way to evaluate the dynamic mechanism of proteins by time-resolved crystallography even if their function would not contain a photoreaction.

Free Research Field

構造生物化学

Academic Significance and Societal Importance of the Research Achievements

ヒトのシトクロムP450はホルモン合成や薬物代謝に深く関与することから、疾病を防ぐための研究が長年にわたって行われてきた。また、植物や細菌など多くの生物にもシトクロムP450は存在し、植物の農薬への耐性や環境汚染物質の浄化の研究などに用いられている。したがって、本課題が実施したP450の活性部位での多様な基質認識や反応メカニズムの研究は、人間にとって有用な化合物生産のための触媒としてシトクロムP450の反応を利用するための研究開発へと発展すると期待される。