Synthesis of natural modified nucleosides and their mass spectrometric quantification in cells and tissue (English shop)

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Natural nucleoside modifications play essential roles in regulating genetic information and maintaining the accuracy of protein synthesis.
Daniel Globisch’s dissertation investigates the chemical synthesis of modified nucleosides and their quantitative determination in biological samples.
The research focuses on developing a mass spectrometry-based method for the parallel analysis of numerous tRNA modifications.
A total of 13 modified nucleosides are synthesized, including six stable isotope-labeled reference compounds.
Quantification is performed using HPLC-ESI-MS with isotope-labeled internal standards.
This analytical approach is applied to investigate tRNA modification profiles in different bacterial species, mammalian tissues, and human cell lines.
The findings reveal significant differences between healthy tissues and cancer cell lines, particularly in the modifications t6A and ms2i6A.
Levels of the mitochondria-specific modification ms2i6A correlate with mitochondrial activity and may serve as a marker for distinguishing healthy and tumor tissues.
Further investigations demonstrate relationships between tRNA modification levels and protein synthesis activity.
Phylogenetic analyses identify characteristic modification patterns among different groups of organisms and reveal changes under environmental stress conditions.
A second major research focus examines the quantification and tissue-specific distribution of the DNA modification 5-hydroxymethylcytosine.
Particularly high concentrations are detected in the central nervous system, with variations between brain regions and age-dependent differences.
Overall, the dissertation establishes analytical foundations for investigating nucleic acid modifications and their significance in gene regulation, metabolism, and disease.