Oral Sessions (Day1, Day2, Day3)
Poster Presentations
- Day 3, May 21（Fri.） Room P1 (Zoom)
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3P-21 PDF
Development of LC-TriVersa NanoMate/HRMS/MS-based analytical platform for the exploration of novel lipid ligands of immunoreceptors
C-type lectin receptors (CLRs) are a superfamily of proteins that recognize a variety of endogenous and exogenous ligands, including carbohydrate, lipid, and protein components of pathogens and self. Elucidation of the structure of unknown ligands for CLRs and their biosynthetic mechanism will be important information for the development of new therapeutic agents for infectious diseases. The aim of this study was to develop an analytical platform for rapidly determining the structure of lipid ligands for the CLRs. Analyses of acid-fast bacillus extracts were performed with a liquid chromatography connected to a quadrupole Orbitrap high-resolution tandem mass spectrometry (Thermo Fisher Scientific) combined with a TriVersa NanoMate (Advion) (LC-TriVersa NanoMate/HRMS/MS). We have constructed an in-house lipidomics database of over 10 million lipids predicted by 108 lipid classes with 34 different fatty acid side chains. The optimized LC- TriVersa NanoMate/HRMS/MS system with in-house lipidomics database enabled us to simultaneously perform structural analysis of lipid molecules based on HRMS/MS information and fractionation into microplates for a reporter assays. In fact, we successfully identified novel lipid ligands for the inhibitory CLR from acid-fast bacillus extracts using LC-TriVersa NanoMate/HRMS/MS analysis and reporter assays. This analytical platform represents a potentially useful tool for a high-throughput and comprehensive exploration of lipid ligands.