Halls, Victoria Simone: Synthesis and Characterisation of wALAD Inhibitors and Novel ABAL Probes. - Bonn, 2016. - Dissertation, Rheinische Friedrich-Wilhelms-Universität Bonn.
Online-Ausgabe in bonndoc: https://nbn-resolving.org/urn:nbn:de:hbz:5n-44946
@phdthesis{handle:20.500.11811/6898,
urn: https://nbn-resolving.org/urn:nbn:de:hbz:5n-44946,
author = {{Victoria Simone Halls}},
title = {Synthesis and Characterisation of wALAD Inhibitors and Novel ABAL Probes},
school = {Rheinische Friedrich-Wilhelms-Universität Bonn},
year = 2016,
month = oct,

note = {The here presented work comprises two projects with highly interdisciplinary topics in the chemical biology field. The development of novel small molecule ALAD inhibitors which might show their potential as novel antifilarial, antiplasmodial and antimicrobial agents, herbicides or chemical biology tools was the main task of the first project (Section A).
wALADin1 is a benzimidazole-based drug-like small molecule that was found by high throughput screening and was identified as a potent species-specific inhibitor for the endobacterial enzyme wALAD with effectiveness in in vitro and ex vivo studies. Several wALADin1 derivatives were synthesised in this work to further characterise the influence on the inhibitory activity of the 3-trifluoromethylbenzyl and 2-[(2 thienylcarbonyl)amino]ethyl group present in the benzimidazole scaffold of wALADin1. These studies revealed the importance of the unaltered 3- trifluoromethylbenzyl-moiety (R2) for the inhibitory activity of the wALADin compounds and the necessity of the 2-[(2-thienylcarbonyl)amino]ethyl-moiety (R1) for species-specificity, although irrelevant for wALAD inhibition. Additionally, the obtained wALADin compounds were tested in cross species structure activity relationship studies and showed contrasting biological results with various ALAD orthologues from plants, bacteria, metazoa and protozoa. Moreover, an antiplasmodial effect on blood stage Plasmodium falciparum was investigated for wALADin1, wALADin2 and wALADin3 that demonstrably does not result from inhibitory activity on the PfALAD orthologue.
The second project comprised the development of novel probes for aptamer-based affinity labelling (ABAL) of proteins, an approach for the rational, proteome-wide identification of proteins that bind to a particular aptamer (Section B).
This is achieved by labelling aptamers with photoreactive probes and cross-linking them to their target protein in a UV light-dependent and highly specific manner. Successful identification requires isolation of pure samples for mass spectrometry analysis, which was not obtained using the initial ABAL probe. To improve the ABAL procedure novel ABAL probes were developed. These probes carry biotin in combination with a chemically cleavable linker or desthiobiotin as purification tags to enable mild elution of the captured complex and besides phenyl azide (PA), 1,2,4,5 tetrafluorophenyl-3-azide (TPA) and 3-phenyl-3-(trifluoromethyl)-3H-diazirine (TPD) were used as potentially more potent photoreactive moieties.
These novel ABAL probes might help to tap the full potential of the ABAL procedure and develop it into a standard procedure for identifying unknown target proteins.},

url = {http://hdl.handle.net/20.500.11811/6898}
}

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