LC-MS system for analysis of proteomes and posttranslational modifications

• Subject Area: Plant Sciences
• Term: Funded in 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 575854334

Research in the Hippler lab focuses on understanding how plant cells respond and adapt to environmental stress. We primarily use the green alga Chlamydomonas reinhardtii as a model system, combining reverse genetics and proteomics to investigate the assembly, function, and regulation of the photosynthetic machinery in oxygenic photosynthesis. Additionally, we study protein N-glycosylation and its role in ciliary motility in Chlamydomonas. Mass spectrometry-based proteomics is central to our research. With our collaborators, we have applied this technology for untargeted (DDA) and targeted (PRM) quantification, posttranslational modification (PTM) analyses—including phosphorylation, N-glycosylation, carboxylation, and oxygenation—and structural studies using chemical crosslinking. Our current LC-MS system has been in near-continuous use for over 10 years and is now out of service due to failed electronics. Repair is no longer economical, making replacement essential. The new instrument should support high-sensitivity, label-free proteome and PTM analyses with accurate quantification and broad dynamic range. It should also allow for high mass range detection (exceeding 10,000 m/z), enabling comprehensive top-down protein analysis and detailed characterization of multiprotein complexes involved in photosynthesis—a central research focus of the GoPMF Forschungsgruppe. The system should be capable of reliably detecting and analyzing large protein assemblies, including those with molecular weights in the several hundred kilodalton range. This capability is crucial for studying the structural organization and dynamics of photosynthetic complexes. Furthermore, the instrument should include ion mobility-assisted separation, which would enhance native mass spectrometry approaches and improve resolution in complex peptide mixtures. This feature would also significantly aid in distinguishing glycopeptides from their non-glycosylated counterparts.The new mass spectrometer would become part of the instrumentation pool of the MS Proteomics Unit Biology of Plants (MSPUB), which supports proteomics research across the Department of Biology at the University of Münster. Its availability will not only sustain the Hippler lab’s research on photosynthetic regulation and ciliary glycosylation but will also benefit a wide range of users working on diverse biological questions involving protein complexes and posttranslational modifications.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: LC-MS-System zur Analyse von Proteomen und posttranslationalen Modifikationen
• Instrumentation Group: 1700 Massenspektrometer
• Applicant Institution: Universität Münster
• Leader: Professor Dr. Michael Hippler