Messenger, Cassie;
(2022)
Investigating the PKM2-dependent regulation of glycolysis by amino acids.
Masters thesis (M.Phil), UCL (University College London).
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Abstract
PKM2 function can be regulated by multiple allosteric ligands and it is not yet well understood how these concurrent ligands come together to illicit one output for PKM2 regulation. Biochemically, amino acids that are PKM2 inhibitors and activators have been used in competition to investigate their combined effect on PKM2 activity, however this has not been monitored in cells including any effects this may have on wider metabolism. In order to measure glucose metabolism and amino acid concentration within the cell, an LC-MS method was optimised in attempt to improve the current methods using a triple quadrupole mass spectrometer, however this method was not robust or reliable enough for requirements. This project confirms that changing media conditions with various alanine to serine ratios, influences the intracellular concentration of alanine and serine. Subsequently, using labelled glucose to track how glucose is broken down in high inhibitor (alanine) and high activator (serine) conditions. Data confirm that there is no significant difference in glycolytic intermediate production between these two conditions. A trend of increased carbons metabolised through the serine production pathway in the high inhibitor treatment condition was observed, however this difference was not significant. This trend suggests there may be a build-up of glycolytic intermediates when PKM2 is inhibited, and therefore a higher production rate of serine. Data also suggest that there is an increased rate of TCA cycle intermediate production when cells are treated with a high activator A:S ratio, suggesting this media condition is influencing an increased activity of PKM2 compared to the high inhibitor treatment condition.
| Type: | Thesis (Masters) |
|---|---|
| Qualification: | M.Phil |
| Title: | Investigating the PKM2-dependent regulation of glycolysis by amino acids |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2022. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10150549 |
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