Development of atherosclerosis, the hardening of the arteries, is dependent on levels of serum cholesterol, which is regulated by the liver via LDL receptors (LDLR). The expression and internalization of LDL receptors depend on several proteins including PCSK9. In fact, previous studies in our laboratory have shown that NEU1 down regulation leads to LDLR hypersialylation which results in its stabilization via reduced interactions with PCSK9. New evidence suggests that NEU1 which de-sialylates LDLR, may affect the ability of another hepatic receptor, the asialoglycoprotein receptor (ASGR), which is comprised of ASGR1 and ASGR2, to interact with LDLR potentially causing its internalization and therefore reduced ability to take up LDL. We investigated how sialidase plays a role in the interaction of ASGR with LDLR. Knockdown and overexpression experiments suggest that NEU1 allows stabilization of LDLR at the cell membrane via ASGR interactions. Treatment of HepG2 cells with monensin which inhibits recycling from the early endosome, unveiled a new truncated ASGR1 isoform potentially lacking its lectin motif. This may be a novel regulatory step in ASGR biosynthesis that warrants further studies. Lysosomal inhibition with chloroquine resulted in concurrent accumulations of NEU1, LDLR and ASGR1, further suggesting these proteins are biosynthetically connected. Our studies revealed a novel isoform of ASGR1 in membrane fractions of HepG2 cell lysates that can associate with NEU1 and LDLR. The impact of NEU1 and ASGR1 on the function and stability of LDLR might lead to new clues for lowering serum cholesterol and reducing atherosclerosis. / Thesis / Master of Science (MSc)
| Identifer | oai:union.ndltd.org:mcmaster.ca/oai:macsphere.mcmaster.ca:11375/25317 |
| Date | January 2019 |
| Creators | Fisher, Kathryn |
| Contributors | Igdoura, Suleiman, Biology |
| Source Sets | McMaster University |
| Language | English |
| Detected Language | English |
| Type | Thesis |
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