Native MS Characterization of RNA-Protein Complexes

In recent decades, the role of ribonucleic acids (RNA) in human disease has undergone a profound paradigm shift. Once considered a mere messenger between DNA and proteins, RNA has evolved into a critical player in a wide range of vital biochemical processes. Thus, besides riboswitches, RNA molecules that play an important role in regulatory processes in bacteria, this research project aims to study RNA-protein complexes with high relevance for human health and their underlying non-covalent interactions. This includes the study of microRNA- protein complexes essential for human gene regulation and the analysis of RNA-protein complexes and protein aggregates associated with the pathology of Alzheimer`s disease. The focus of the proposed research project is the development and application of innovative and novel mass spectrometry techniques. These methods will allow the analysis of RNA molecules and RNA-protein complexes in their native state without extensive sample preparation. By combining specialized native nano-electrospray ionization (nano-ESI), top- down mass spectrometry and charge detection mass spectrometry (CDMS), insights will be gained into binding sites, recognition principles, stoichiometry, stability and folding dynamics of RNA molecules and RNA-protein complexes up to the gigadalton range. The proposed research project will provide new insights into highly intricate biochemical processes involving the studied RNA-protein complexes. This will contribute to a better understanding of still poorly understood disease pathogenesis, such as cancer and Alzheimer`s disease, and will be crucial for the development of new RNA-based therapeutic approaches.