{"id":{"repo_id":"uwo","oai_identifier":"oai:uwo.scholaris.ca:20.500.14721/37577"},"canonical_url":"https://search.dev.ndltd.org/etd/uwo/oai:uwo.scholaris.ca:20.500.14721/37577","repository":{"repo_id":"uwo","name":"Western University","base_url":"https://uwo.scholaris.ca/server/oai/request"},"display":{"title":"The Effects of S-glutathionylation on the Structure and Function of the Mitochondrial Calcium Uniporter (MCU)","abstract":"Most calcium (Ca2+) entry into the mitochondrial matrix is regulated by the mitochondrial calcium uniporter (MCU). The amino (N)-terminal domain (NTD) of MCU is a regulatory component of the channel. S-Glutathionylation of Cys97 on the MCU-NTD leads to robust MCU activation and increased matrix Ca2+. Here, I characterized the biophysical and structural changes induced by Cys97 S-glutathionylation by applying optical spectroscopy, light scattering, solution nuclear magnetic resonance (NMR) and live cell functional experiments. S-Glutathionylation increased solvent exposed hydrophobicity, destabilized and caused large structural perturbations in the MCU-NTD. An S-glutathiomimetic mutation was able to closely recapitulate these biophysical and structural effects, but in the absence of oxidative stress. Indeed, HeLa cells expressing MCU with the S-glutathiomimetic mutation, showed increased mitochondrial Ca2+ uptake compared to wild-type MCU expressing cells. Thus, my research revealed new insights into the impact of S-glutathionylation on MCU-NTD and identified the S-glutathiomimetic mutation as a valuable research tool.","abstract_html":"Most calcium (Ca2+) entry into the mitochondrial matrix is regulated by the mitochondrial calcium uniporter (MCU). The amino (N)-terminal domain (NTD) of MCU is a regulatory component of the channel. S-Glutathionylation of Cys97 on the MCU-NTD leads to robust MCU activation and increased matrix Ca2+. Here, I characterized the biophysical and structural changes induced by Cys97 S-glutathionylation by applying optical spectroscopy, light scattering, solution nuclear magnetic resonance (NMR) and live cell functional experiments. S-Glutathionylation increased solvent exposed hydrophobicity, destabilized and caused large structural perturbations in the MCU-NTD. An S-glutathiomimetic mutation was able to closely recapitulate these biophysical and structural effects, but in the absence of oxidative stress. Indeed, HeLa cells expressing MCU with the S-glutathiomimetic mutation, showed increased mitochondrial Ca2+ uptake compared to wild-type MCU expressing cells. Thus, my research revealed new insights into the impact of S-glutathionylation on MCU-NTD and identified the S-glutathiomimetic mutation as a valuable research tool.","abstract_has_math":false,"creators":["Pawakian, Arine"],"institution":"The University of Western Ontario","degree_name":"M Sc","degree_level":null,"degree_discipline":"Physiology and Pharmacology","degree_department":null,"school":null,"contributors":[],"advisors":["Stathopulos, Peter B."],"committee_chairs":[],"committee_members":[],"year":2024,"date_issued":"2024-01-25","date_published":"2024-01-25","updated_at":"2026-07-27T21:56:09Z","subjects":["calcium signaling","mitochondrial bioenergetics","mitochondrial calcium uptake","oxidative stress","post-translational modifications","protein structure","reactive oxygen species","S-glutathiomimetic","S-gluthathionylation"],"languages":["en_ca"],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://hdl.handle.net/20.500.14721/37577","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Stathopulos, Peter B."]},{"key":"dc:creator","label":"Author","values":["Pawakian, Arine"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2025-07-10T21:38:21Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2025-07-10T21:38:21Z"]},{"key":"dc:date.issued","label":"Date","values":["2024-01-25"]},{"key":"dc:publisher","label":"Institution","values":["The University of Western Ontario"]},{"key":"dc:type","label":"Dc Type","values":["thesis"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Physiology and Pharmacology"]},{"key":"thesis:degree_name","label":"Degree Name","values":["M Sc"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["calcium signaling","mitochondrial bioenergetics","mitochondrial calcium uptake","oxidative stress","post-translational modifications","protein structure","reactive oxygen species","S-glutathiomimetic","S-gluthathionylation"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en_ca"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://hdl.handle.net/20.500.14721/37577"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The thesis cover page in the PDF document includes references to Western University’s previous institutional repository platform, known as Scholarship@Western, and links to that platform (beginning with ir.lib.uwo.ca). In citing or referring to this thesis, use the DOI or handle from this page instead. Sample citation: Author name, \"Thesis title.\" (Year). Western University Open Repository. https://doi.org/10.71858/123456."]},{"key":"dc:description.abstract","label":"Abstract","values":["Most calcium (Ca2+) entry into the mitochondrial matrix is regulated by the mitochondrial calcium uniporter (MCU). The amino (N)-terminal domain (NTD) of MCU is a regulatory component of the channel. S-Glutathionylation of Cys97 on the MCU-NTD leads to robust MCU activation and increased matrix Ca2+. Here, I characterized the biophysical and structural changes induced by Cys97 S-glutathionylation by applying optical spectroscopy, light scattering, solution nuclear magnetic resonance (NMR) and live cell functional experiments. S-Glutathionylation increased solvent exposed hydrophobicity, destabilized and caused large structural perturbations in the MCU-NTD. An S-glutathiomimetic mutation was able to closely recapitulate these biophysical and structural effects, but in the absence of oxidative stress. Indeed, HeLa cells expressing MCU with the S-glutathiomimetic mutation, showed increased mitochondrial Ca2+ uptake compared to wild-type MCU expressing cells. Thus, my research revealed new insights into the impact of S-glutathionylation on MCU-NTD and identified the S-glutathiomimetic mutation as a valuable research tool."]},{"key":"dc:title","label":"Title","values":["The Effects of S-glutathionylation on the Structure and Function of the Mitochondrial Calcium Uniporter (MCU)"]}]}],"canonical_facts":{"dc:contributor.advisor":["Stathopulos, Peter B."],"dc:creator":["Pawakian, Arine"],"dc:date.accessioned":["2025-07-10T21:38:21Z"],"dc:date.available":["2025-07-10T21:38:21Z"],"dc:date.issued":["2024-01-25"],"dc:description":["The thesis cover page in the PDF document includes references to Western University’s previous institutional repository platform, known as Scholarship@Western, and links to that platform (beginning with ir.lib.uwo.ca). In citing or referring to this thesis, use the DOI or handle from this page instead. Sample citation: Author name, \"Thesis title.\" (Year). Western University Open Repository. https://doi.org/10.71858/123456."],"dc:description.abstract":["Most calcium (Ca2+) entry into the mitochondrial matrix is regulated by the mitochondrial calcium uniporter (MCU). The amino (N)-terminal domain (NTD) of MCU is a regulatory component of the channel. S-Glutathionylation of Cys97 on the MCU-NTD leads to robust MCU activation and increased matrix Ca2+. Here, I characterized the biophysical and structural changes induced by Cys97 S-glutathionylation by applying optical spectroscopy, light scattering, solution nuclear magnetic resonance (NMR) and live cell functional experiments. S-Glutathionylation increased solvent exposed hydrophobicity, destabilized and caused large structural perturbations in the MCU-NTD. An S-glutathiomimetic mutation was able to closely recapitulate these biophysical and structural effects, but in the absence of oxidative stress. Indeed, HeLa cells expressing MCU with the S-glutathiomimetic mutation, showed increased mitochondrial Ca2+ uptake compared to wild-type MCU expressing cells. Thus, my research revealed new insights into the impact of S-glutathionylation on MCU-NTD and identified the S-glutathiomimetic mutation as a valuable research tool."],"dc:identifier.uri":["https://hdl.handle.net/20.500.14721/37577"],"dc:language.iso":["en_ca"],"dc:publisher":["The University of Western Ontario"],"dc:subject":["calcium signaling","mitochondrial bioenergetics","mitochondrial calcium uptake","oxidative stress","post-translational modifications","protein structure","reactive oxygen species","S-glutathiomimetic","S-gluthathionylation"],"dc:title":["The Effects of S-glutathionylation on the Structure and Function of the Mitochondrial Calcium Uniporter (MCU)"],"dc:type":["thesis"],"thesis:degree_discipline":["Physiology and Pharmacology"],"thesis:degree_name":["M Sc"]},"updated_at":"2026-07-27T21:56:09Z"}