dc.rights.license | CC-BY-NC-ND | |
dc.contributor.advisor | Baggelaar, Marc | |
dc.contributor.author | Stokkel, Marijke | |
dc.date.accessioned | 2022-01-18T00:00:18Z | |
dc.date.available | 2022-01-18T00:00:18Z | |
dc.date.issued | 2022 | |
dc.identifier.uri | https://studenttheses.uu.nl/handle/20.500.12932/372 | |
dc.description.abstract | S-Palmitoylation is a post translational modification (PTM) associated with numerous cancers and
neurological diseases. It is a reversible type of lipidation defined by the addition of a palmitic acid to a
cysteine residue. Regulation of this PTM depends on the activity of palmitoyl S-acyl transferases and
palmitoyl thioesterases which add and remove the palmitoyl moiety, respectively. S-palmitoylation can
have various effects on proteins, of which a change in intracellular localization is most substantiated.
While methods for large scale identification of palmitoylated proteins have become well established, the
characterization of S-palmitoylation dependent translocations has so far only been studied on a single
protein basis. This study aimed to identify S-palmitoylation dependent translocating proteins. The broad
spectrum depalmitoylase inhibitor Palmostatin B (PalmB) was employed in combination with differential
centrifugation based spatial proteomics to determine translocating proteins. In addition, SILAC-TMT
hyperplexing was used to allow for the combined quantitation of treatment groups and spatial
proteomics fractions. Using this method we identified 48 potentially translocating proteins. A high
overrepresentation of S-palmitoylated proteins was observed among the translocators indicating that
the translocation of these enriched proteins may have occurred in response to a change in
palmitoylation. Additionally, a STRING database search found many interacting proteins within this
group suggesting that some translocations may occur as a result of the translocation of a binding
partner. We demonstrate that spatial proteomics can be used as a hypothesis-generating method to
identify potential S-palmitoylation dependent translocating proteins. These potential translocators can
be used as a starting point for further study into S-palmitoylation dependent protein localization. | |
dc.description.sponsorship | Utrecht University | |
dc.language.iso | EN | |
dc.subject | Determining proteins that change subcellular localization upon treatment with the depalmitoylase inhibitor Palmostatin B. | |
dc.title | Characterization of S-palmitoylation dependent protein localization by hyperplexed spatial proteomics | |
dc.type.content | Master Thesis | |
dc.rights.accessrights | Open Access | |
dc.subject.keywords | S-palmitoylation, Palmostatin B, spatial proteomics, translocation, SILAC-TMT, higher order multiplexing | |
dc.subject.courseuu | Molecular and Cellular Life Sciences | |
dc.thesis.id | 1741 | |