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Visualizing the connection between L-arginine metabolism and the TCA cycle in Mycobacterium tuberculosis infection in primary mouse macrophages

Robillard, Michelle
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1592168838162627

Abstract Details

2020, MS, University of Cincinnati, Medicine: Immunology.
Select enzymes assemble in complexes forming catalytic networks that channel metabolic substrates. This channeling can allow utilization of metabolites in low concentrations and serve to shield substrates from competing enzymes. The L-arginine regeneration cycle is an example of this type of multi-enzymatic complex that we hypothesize locates to the mitochondrion to enhance substrates and provide products integral to the tricarboxylic acid cycle. Still, enzymatic complexes can be difficult to verify because of their instability in in vitro environments. Optimizing fluorescence staining for confocal microscopy can be performed to enhance images for peak visualization of proteins or complexes within a cell. Visualizing protein-protein, or protein-organelle, co-localization is a useful technique making use of confocal imaging, which can provide support for interaction between two or more targets. By using this technique, the intercellular environment is preserved and protein complexes – or those interacting with organelles – can be visualized in situ. In the current project, we aim to visualize an enzyme involved in the L-arginine regeneration cycle – argininosuccinate synthase 1 (ASS1) – and the mitochondria. We expect these to co-localize, which will be a first step in understanding how key metabolites are shuttled between the mitochondria and the L-arginine regeneration machinery. We plan to evaluate other metabolic networks in the future using techniques we are currently developing. Understanding the intracellular movement of metabolites within enzyme complexes and other intracellular compartments is expected to uncover novel regulatory checkpoints, which may prove fruitful in designing future therapeutic strategies.
Joseph Qualls, Ph.D. (Committee Chair)
George Deepe, M.D. (Committee Member)
Jonathan Katz, Ph.D. (Committee Member)
57 p.
Immunology
Metabolism; L-arginine; TCA cycle; Confocal Microscopy; Enzyme Complexes; Mycobacterium tuberculosis

Recommended Citations

Citations

  • Robillard, M. (2020). Visualizing the connection between L-arginine metabolism and the TCA cycle in Mycobacterium tuberculosis infection in primary mouse macrophages [Master's thesis, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1592168838162627

    APA Style (7th edition)

  • Robillard, Michelle. Visualizing the connection between L-arginine metabolism and the TCA cycle in Mycobacterium tuberculosis infection in primary mouse macrophages. 2020. University of Cincinnati, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1592168838162627.

    MLA Style (8th edition)

  • Robillard, Michelle. "Visualizing the connection between L-arginine metabolism and the TCA cycle in Mycobacterium tuberculosis infection in primary mouse macrophages." Master's thesis, University of Cincinnati, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1592168838162627

    Chicago Manual of Style (17th edition)

ucin1592168838162627
241
© 2020, some rights reserved.Creative Commons License Visualizing the connection between L-arginine metabolism and the TCA cycle in Mycobacterium tuberculosis infection in primary mouse macrophages by Michelle Robillard is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by University of Cincinnati and OhioLINK.