Grossenbacher, Philipp (2022). Development of Synthetic Research Tools for Structural and Functional Studies of Mitochondrial Carriers and Other Membrane Proteins. (Thesis). Universität Bern, Bern
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22grossenbacher_p.pdf - Thesis Available under License Creative Commons: Attribution-Noncommercial (CC-BY-NC 4.0). Download (29MB) | Preview |
Abstract
Calcium signalling mediates many physiological functions such as regulating cell death/ survival, cell proliferation/ differentiation and gene expression. The calcium signals within cells originate in a release of bivalent calcium cations (Ca2+) from the endoplasmic reticulum (ER), which is the main store of intracellular calcium. The endoplasmic reticulum is a highly dynamic structure within the cell and can tether itself to the plasma membrane or membranes of other organelles in the cell such as the mitochondrion. Even though it is already established that a rise in the intracellular Ca2+ concentration signals an increased energy demand, the many relationships between the calcium concentration and the energy metabolism remain unknown. With the aim to reveal individual players in the calcium regulation pathway and gain insight into their function, different strategies were envisioned as part of a SNF funded Sinergia project, which involves multiple project partners. Two of the envisioned chemical biology strategies involved synthesis of chemical tools. The first strategy required fluorescent antibodies against targets of the calcium signalling/communication machinery for fluorescence microscopy and western blot applications. The second strategy entailed ligands to stabilise the conformation of a Ca2+-controlled mitochondrial solute carrier (mSLC), in order to solve the structure of this particular mSLC by cryo-EM. We designed and synthesised compounds that allow for easy and functional modification of antibodies, rendering them fluorescent whilst remaining functional for fluorescence microscopy and western blot experiments. Furthermore, we have developed synthetic ligands that stabilise a particular conformation of the highly dynamic mSLC Citrin (AGC2, SLC25A13) to aid determination of its structure by cryo-EM. In several parallel projects, various tool compounds and probes for biochemical investigations were synthesised and biologically evaluated. This comprises inhibitors of the parasite Echinococcus multilocularis, pH-dependent fluorescent probes for proteoliposomes and giant unilamellar vesicle (GUV) applications, trifunctional bioorthogonal probes for proteomic and target identification studies, store-operated calcium entry (SOCE) inhibitors and positive allosteric modulators of the G protein-coupled adenosine A1 receptor.
| Item Type: | Thesis |
|---|---|
| Dissertation Type: | Cumulative |
| Date of Defense: | 16 December 2022 |
| Subjects: | 500 Science > 540 Chemistry 500 Science > 570 Life sciences; biology 600 Technology > 610 Medicine & health |
| Institute / Center: | 04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Biochemistry and Molecular Medicine |
| Depositing User: | Hammer Igor |
| Date Deposited: | 07 Jan 2025 09:56 |
| Last Modified: | 07 Jan 2025 23:25 |
| URI: | https://boristheses.unibe.ch/id/eprint/5717 |
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