Coenzyme engineering for the study of RNA methyltransferase

2019 Fiscal Year Annual Research Report

• Project/Area Number: 18K14656
• Research Institution: Okinawa Institute of Science and Technology Graduate University
• Principal Investigator: LAURINO Paola 沖縄科学技術大学院大学, タンパク質工学・進化ユニット, 准教授 (90812256)
• Project Period (FY): 2018-04-01 – 2020-03-31
• Keywords: SAM analogues / methionine / adenosyltransferase / Methyltransferases / kinetic assays

Outline of Annual Research Achievements

During these last two years we have:
1. studied the methionine adenosyl transferase and exploited the catalytic promiscuity of this enzyme to synthetize biochemically synthetic analogues of SAM. 2. Establish a HPLC method to detect the formation of the SAM analogues and purify them. 3. Study three different bacterial Methyltransferases scaffold as potential starting point for our engineering study. 4. We established a UPLC-MS method to run kinetics assays for the MTases. 5. We defined the kinetics parameters for the synthetic cofactors

Overall the data collected in this period of time allowed us to write a manuscript (in preparation) and collect enough data for a second one.