Identification of NBD-labeled peptides by using photo irradiation-induced NBD loss

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K01956
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biomolecular chemistry
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Asanuma Miwako 国立研究開発法人理化学研究所, 環境資源科学研究センター, 技師 (70381598)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: 質量分析 / 標的タンパク質 / 蛍光 / 標識 / ケミカルバイオロジー

Outline of Final Research Achievements

We have previously made it possible to identify nitrobenzoxadiazole (NBD)-labeled proteins using a turn-ON fluorescent affinity labeling method and mass spectrometry. However, it was not easy to identify NBD-labeled peptides and determine the NBD-modified sites by mass spectrometry. In this study, in order to identify the NBD-modified sites of NBD-labeled proteins, we tried to establish a method for selective detection of NBD-labeled peptides using liquid chromatography-mass spectrometry, focusing on a phenomenon of the loss of NBD moiety from NBD-labeled peptide by photo-irradiation, and also the sample preparation method for the MS analysis. As a result, we established a method to recover NBD-labeled peptides and succeeded in identification of some of the NBD-labeled peptides and the NBD-modified sites.

Free Research Field

質量分析

Academic Significance and Societal Importance of the Research Achievements

本研究により、NBD標識されたタンパク質のNBD修飾部位を同定することが可能となった。今後、未だ作用機序の解明されていない様々な生物活性物質に本手法を活用することで、生物活性化合物の標的タンパク質や周辺タンパク質の探索が進み、医薬品開発や分子生物学的分野への展開が期待される。