Native metabolomics for mass spectrometry-based siderophore discovery.

Area of Science:

• Metabolomics
• Analytical Chemistry
• Biochemistry

Background:

• Organisms utilize specialized pathways involving siderophores and metallophores for metal acquisition.
• Identifying these metal-binding molecules is essential for understanding biological processes.

Purpose of the Study:

• To present a novel mass spectrometry-based approach, Native Metabolomics, for discovering metal-binding molecules.
• To detail the workflow of Native Metabolomics, integrating liquid chromatography, pH adjustment, metal infusion, and ion identity molecular networking (IIMN).

Main Methods:

• Native Metabolomics employs liquid chromatography-mass spectrometry (LC-MS) with post-chromatographic pH adjustment and metal infusion.
• Ion identity molecular networking (IIMN) analyzes mass-to-charge ratio (m/z) offsets, chromatographic profiles, and fragmentation patterns to identify metal-binding species.
• The method is applicable to diverse sample types, including microbial supernatants, biological tissues, and environmental samples.

Main Results:

• This approach has successfully identified novel metal-binding metabolites.
• It revealed yersiniabactin as a zincophore, lepotchelin natural products as cyanobacterial metallophores, and iron-binding antioxidants in traditional medicine.

Conclusions:

• Native Metabolomics is a versatile and powerful strategy for elucidating biological metal-binding molecules.
• The method can be adapted to any LC-MS system for simultaneous analysis of multiple metals, offering broad applicability.