Plasma membrane (PM) proteins are at the interface between the cell and the external environment and are therefore the most accessible to therapeutic drugs. I utilized cationic silica beads and mass spectrometry (MS)-based proteomics to enrich for PM proteins of human cardiomyocytes, coronary smooth muscle cells, and coronary endothelial cells. The enrichment of PM proteins was confirmed and 1006 proteins were specifically filtered and enriched into a set of known and novel cardiomyocyte PM-associated proteins of which 42% had PM-associated gene ontology annotations and/or predicted transmembrane helices. Two novel candidates, namely popeye domain-containing protein 2 (POPDC2) and protein kinase C and casein kinase substrate in neurons protein 3 (PACSIN3) were selected and found to have confirmed PM localization. In conclusion, silica bead membrane extraction combined with MS-based proteomics successfully enriched for PM proteins of the human heart of which two novel candidate proteins were shown to have confirmed PM localization.
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/25880 |
| Date | 13 January 2011 |
| Creators | Noronha, Melissa |
| Contributors | Gramolini, Anthony |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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