Rapid MALDI-MS/MS-Based Profiling of Lipid A Species from Gram-Negative Bacteria Utilizing Trapped Ion Mobility Spectrometry and mzmine

Rudt E.; Froning M.; Heuckeroth S.; Ortmann L.; Diemand J.; Hörnschemeyer L.; Pleger A.; Vinzelberg M.; Schmid R.; Pluskal T.; Dobrindt U.; Hayen H.; Korf A.

Forschungsartikel (Zeitschrift) | Peer reviewed

Zusammenfassung

Lipid A, a crucial component of lipopolysaccharides (LPS), plays a pivotal role in the pathogenesis of Gram-negative bacteria. Lipid A patterns are recognized by mammals and can induce immunostimulatory effects. However, the outcome of the interaction is highly dependent on the chemical composition of individual lipid A species. The diversity of potential fatty acyl and polar headgroup combinations in this complex saccharolipid presents a significant analytical challenge. Current mass spectrometry (MS)-based lipid A methods are focused on either direct matrix-assisted laser desorption/ionization (MALDI)-MS screening or comprehensive structural elucidation by tandem mass spectrometry (MS/MS) hyphenated with separation techniques. In this study, we developed an alternative workflow for rapid lipid A profiling covering the entire analysis pipeline from sample preparation to data analysis. This workflow is based on microextraction and subsequent MALDI-MS/MS analysis of uropathogenic Escherichia coli utilizing trapped ion mobility spectrometry (TIMS), followed by mzmine data processing. The additional TIMS dimension served for enhanced sensitivity, selectivity, and structural elucidation through mobility-resolved fragmentation via parallel accumulation-serial fragmentation (PASEF) in parallel reaction monitoring (prm)-mode. Furthermore, mzmine enabled automated MS/MS acquisition by adapting the spatial ion mobility-scheduled exhaustive fragmentation (SIMSEF) strategy for MALDI spot analysis. It also facilitated robust lipid A annotation through a newly developed extension of the rule-based lipid annotation module, allowing for the custom generation of lipid classes, including specific fragmentation rules. In this study, the first publication of lipid A species’ collision cross section (CCS) values is reported, which will enhance high-confidence lipid A annotation in future studies.

Details zur Publikation

FachzeitschriftAnalytical Chemistry (Anal Chem)
Jahrgang / Bandnr. / Volume97
Ausgabe / Heftnr. / Issue14
Seitenbereich7781-7788
StatusVeröffentlicht
Veröffentlichungsjahr2025 (15.04.2025)
Sprache, in der die Publikation verfasst istEnglisch
DOI10.1021/acs.analchem.4c05989
Link zum Volltexthttps://api.elsevier.com/content/abstract/scopus_id/105002650424
StichwörterLipid A; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tandem Mass Spectrometry; Ion Mobility Spectrometry; Uropathogenic Escherichia coli; Gram-Negative Bacteria

Autor*innen der Universität Münster

Dobrindt, Ulrich
Institut für Hygiene
Froning, Matti
Professur für Analytische Chemie (Prof. Hayen)
Hayen, Heiko
Professur für Analytische Chemie (Prof. Hayen)
Heuckeroth, Steffen
Professur für Analytische Chemie (Prof. Karst)
Ortmann, Lucas
Professur für Analytische Chemie (Prof. Hayen)
Rudt, Edward
Professur für Analytische Chemie (Prof. Hayen)