Proteins rarely act alone. They form complexes, carry chemical modifications, and exist as multiple distinct proteoforms that together determine cellular function. Understanding this molecular complexity is a prerequisite for advancing personalized medicine, yet the tools to characterize it at sufficient resolution have only recently become available.
Modern advances enable treatments increasingly tailored to an individual's molecular profile through greater precision in diagnosis, therapy, and prognosis. This approach aims to address disease causes rather than symptoms, with the potential for more targeted treatments with fewer side effects compared to conventional approaches. Achieving this requires analytical methods capable of resolving the full chemical heterogeneity of disease-relevant proteins, including modification states and proteoform distributions that previously remained inaccessible to conventional approaches.
This project focused on the 5'-adenosine monophosphate-activated protein kinase (AMPK), known as the guardian of cellular energy, a central regulator of cellular energy metabolism operating through complex phosphorylation networks, and associated with metabolic disorders, cardiovascular diseases, and viral infections. Native top-down mass spectrometry was applied as the core methodology, allowing intact protein complexes to be analyzed while preserving non-covalent interactions and capturing the full proteoform landscape.
The project brought together three complementary partners. The outgoing phase was carried out at the University of Wisconsin-Madison, chosen for its strong expertise in top-down proteomics, cardiac research, and kinase biology. The return phase was hosted at the University of Lübeck and CSSB Hamburg, providing a research environment centered on structural biology and native mass spectrometry. The non-academic placement at Bruker Daltonics connected the academic research directly to mass spectrometry instrument development, enabling knowledge transfer in both directions between fundamental research and industrial application.
The broader objective was to establish a methodology applicable beyond AMPK, contributing to the molecular characterization of disease-relevant kinase complexes and illustrating how fundamental research in structural proteomics can inform future clinical development.
