Development of the binding position analysis of target biomolecules by using photoaffinity probes and LA-LDI MS

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K12753
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Chemical biology
• Research Institution: University of Tsukuba
• Principal Investigator: KITA Masaki 筑波大学, 数理物質系, 准教授 (30335012)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 天然物ケミカルバイオロジー / 標的分子同定 / 質量分析法 / 光親和性プローブ / LA-LDI MS

Outline of Final Research Achievements

To establish new binding mode analysis methods of target biomacromolecules, we aimed to develop photoaffinity probes that can covalently bind to target molecules and efficiently analyze their fragment peptides by using label-assisted laser desorption/ionization mass spectrometry (LA-LDI MS). We found that 6-amidopyrene (apy) derivatives were highly detectable by the LDI MS instrument. An apy-conjugated photoaffinity derivative of an antitumor natural product, aplyronine A, was then synthesized. By the irradiation of UV light (365 nm), this probe efficiently photoreacted with solvent molecules and form covalent bonds. Furthermore, actin was quantitatively labeled with an apy NHS ester probe of aplyronine A, and the sequence of an apy-labeled peptide was established by MS/MS analysis.

Free Research Field

天然物有機化学、ケミカルバイオロジー