- Title
- Evaluation of model systems for the study of protein association / incorporation of Beta-Methylamino-L-Alanine (BMAA)
- Creator
- Visser, Claire
- Subject
- Neurotoxic agents
- Subject
- Nervous system -- Diseases
- Date Issued
- 2011
- Date
- 2011
- Type
- Thesis
- Type
- Masters
- Type
- MSc
- Identifier
- vital:10314
- Identifier
- http://hdl.handle.net/10948/1451
- Identifier
- Neurotoxic agents
- Identifier
- Nervous system -- Diseases
- Description
- β-methylamino-L-alanine (BMAA) is thought to be a contributing factor of Amyotrophic Lateral Sclerosis-Parkinsonism Dementia Complex (ALS/PDC). It has been shown that the levels of toxin ingestion by humans are too low to cause disease. However, it has recently been theorized that this toxin is bioaccumulated within cells. Via a process of slow release from this reservoir, the BMAA is able to bring about neurotoxicity. Mechanisms of uptake and bioaccumulation of BMAA have been proposed in several publications; however the mechanism of protein incorporation of BMAA has not yet been identified. Identifying suitable model systems would be a prerequisite in order for future studies on BMAA protein incorporation. Three specific models were therefore chosen for investigation; mammalian cell lines including C2C12 and HT29, a prokaryotic (E. coli) expression system and yeast cells. The cytotoxity of BMAA was established for the mammalian cell lines and further investigation of BMAA incorporation into cellular proteins was performed on all three above mentioned models. Samples were run on HPLC-MS in order to determine uptake of BMAA into cells or lack thereof. Results indicate negligible cytotoxicity as measured by MTT and CellTitre Blue assays, limited uptake and protein incorporation of BMAA within the prokaryotic model and insignificant uptake of BMAA by yeast cells. Although the uptake of BMAA in the prokaryotic model was not extensive, there was indeed uptake. BMAA was not only taken up into the cells but was also observed in inclusion body protein samples after hydrolysis. After further investigation and use, this model could very well provide researchers with information pertaining to the mechanism of association of BMAA with proteins. Although the other models provided negative results, this research was valuable in the sense that one can narrow down the number of possible model systems available. Also, in seeking models for studying protein association/incorporation, the use of the final target cell is not relevant or necessary as the purpose of the research was to identify a model system in which the mechanism of protein association/incorporation can, in future, be studied.
- Format
- viii, 71 leaves
- Format
- Publisher
- Nelson Mandela Metropolitan University
- Publisher
- Faculty of Science
- Language
- English
- Rights
- Nelson Mandela Metropolitan University
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