Development of Alkyne-tag Technologies for Identification of Protein Adducts

Abstract

Alkyne is a tiny functional group and stable in cells. In addition, alkyne has a strong Raman signal in the cellular silent region, where endogenous biomolecules do not show Raman signals. Taking advantage of these properties, we developed the Alkyne-tag Raman Imaging (ATRI) method, in which alkyne is introduced as a tag into a small molecule and its subcellular localization is visualized by Raman microscope. We also applied ATRI method for the analysis of protein adducts and investigated a method to detect peptides labeled with alkyne-tagged compound. As a result, we succeeded in developing Alkyne-tag Raman Screening (ATRaS) method by using Surface Enhanced Raman Scattering (SERS) on silver nanoparticles. In addition, we have recently developed Alkyne-tag enrichment on Silver-immobilized Solid support (ASiS) method, a technique to capture and enrich alkyne-modified molecules by using the interaction between silver and alkyne. Furthermore, we combined ASiS method and ATRaS method to identify protein adducts. In this presentation, we will introduce these three alkyne-tag technologies and show the results of applying ASiS-ATRaS method to analyze the protein acylation.