The research presented in the thesis describes the development and implementation of solution based NMR methods that provide 3D structural information on the protein-small molecule complexes. These... Show moreThe research presented in the thesis describes the development and implementation of solution based NMR methods that provide 3D structural information on the protein-small molecule complexes. These methods can be critical for structure based drug design and can be readily applied in the early stages of preclinical drug discovery and fragment based drug discovery Show less
Urinary tract infection is a complex clinical entity a common infectious disease that encompasses a variety of clinical syndromes with a positive bacterial culture as common denominator. This... Show moreUrinary tract infection is a complex clinical entity a common infectious disease that encompasses a variety of clinical syndromes with a positive bacterial culture as common denominator. This thesis provides an exhaustive exploratory study of the metabolic pattern of patients affected by urinary tract infection and Here this complex clinical entity was investigated with a multiplatform approach. Each of the used platforms added a unique perspective to the further understanding of the infection process. The assessment of the bacterial growth (NMR), of the host response (LC__MS) and of the physiological status (GC-APCI-MS) could eventually be useful during the assessment of the disease severity and/or decision making Show less
The thesis aims at combining metabolomics with other methods to investigate the regulation of the TIA biosynthesis and how this is connected with other pathways and the plant__s physiology and... Show moreThe thesis aims at combining metabolomics with other methods to investigate the regulation of the TIA biosynthesis and how this is connected with other pathways and the plant__s physiology and development. It reviews the biosynthesis studies of Catharanthus roseus. An HPLC method is described for both precursors and alkaloids which was validated to analyze alkaloid contents in different organs, its relationship with leaf age and growth of C. roseus plants. A metabolomics investigation of C. roseus varieties with different flower colors by NMR profiling method is also reported. Besides, it explores the effect of JA/MeJA treatment on TIA biosynthesis in different organs and the influence of flowering on TIA levels in C. roseus plants. Overexpression of ORCA3 and G10H in C. roseus plants is reported and the effect on alkaloid biosynthesis and general metabolism is revealed by NMR-metabolomics. Finally, it reports on C. roseus plants fed with 13C-labeled glucose to study the flux into different metabolic pathways, and gives a summary and discussion about further perspectives. Show less
By utilizing paramagnetic NMR techniques, the structure and dynamics of the P450cam system were investigated. The analysis of PCS and RDC illuminated the stereo-specific final complex of Pdx and... Show moreBy utilizing paramagnetic NMR techniques, the structure and dynamics of the P450cam system were investigated. The analysis of PCS and RDC illuminated the stereo-specific final complex of Pdx and P450cam, while the results of PRE demonstrated the presence of a transient encounter complex. Furthermore, the significant insights of the interaction in the interface were uncovered by X-ray crystallography. Currently, the nature of Pdx effector activity is under debate. Since paramagnetic NMR experiments are applicable to solution studies at ambient temperature, PCS, RDC and PRE methods can further resolve the molecular mechanism of P450cam in the future. Show less
