multiple identification of non-ATP-dependent kinases based on registered crystal structures of enzymes with unknown function

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K22385
• 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: Osaka Medical and Pharmaceutical University (2021); Kyoto University (2019-2020)
• Principal Investigator: Fujihashi Masahiro 大阪医科薬科大学, 医学部, 准教授 (10397581)
• Project Period (FY): 2019-06-28 – 2022-03-31
• Keywords: ピロリン酸 / 結晶構造

Outline of Final Research Achievements

We challenged to remove the common notion that “The phosphate group donor in biological process is ATP". We searched kinases that have no pocket to recognize ATP from kinases registered in the three-dimensional structure database. Their pyrophosphate-dependent kinase (PPi-kinase) activities were investigated against the ligand library for potential phosphate acceptors. Several PPi-kinase were identified by this strategy. Also, we transplanted the myo-inositol recognition residues of ATP-dependent myo-inositol-3-kinase into PPi-dependent myo-inositol-1-kinase based on their three-dimensional structure similarity. The transplantation successfully converted PPi-dependent myo-inositol-1-kinase into PPi-dependent myo-inositol-3-kinase. Some phosphorylated products are industrially important. Our results may contribute to develop the phosphorylation process that is friendly to both the environment and our wallet.

Free Research Field

構造生物化学

Academic Significance and Societal Importance of the Research Achievements

リン酸化反応は生体内における様々な場面で重要な役割を果たしている。これまで「生体内リン酸化反応のリン酸基供与体は ATP」と信じられてきたが、本研究により広く例外が存在する可能性が示された。今後ATPを使わない様々なリン酸化反応が発見され、生命活動の仕組みがより深く理解されることが期待される。
またリン酸化は、化粧品の有効成分の安定性を高めるなど、産業的にも重要である。本研究で成功したPPi-kinaseの創成手法を応用すれば、ATPに比べて3桁安価なピロリン酸を用いて様々な物質を酵素でリン酸化することが可能になり、環境的にも経済的にも優しい有用物質生産の実現が期待される。