Saucedo-Rodriguez 2023 MiP2023: Difference between revisions
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{{Abstract | {{Abstract | ||
|title=[[Image:MiPsocietyLOGO.JPG|left|100px|Mitochondrial Physiology Society|MiPsociety]] Effect of succinate dehydrogenase deficiency on mitochondrial function. | |title=[[Image:MiPsocietyLOGO.JPG|left|100px|Mitochondrial Physiology Society|MiPsociety]] Effect of succinate dehydrogenase deficiency on mitochondrial function. | ||
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|year=2023 | |year=2023 | ||
|event=MiP2023 Obergurgl AT | |event=MiP2023 Obergurgl AT | ||
|abstract=Introduction: Succinate dehydrogenase (SDH) connects the TCA cycle by oxidizing succinate to fumarate and the respiratory chain by transferring electrons to ubiquinone. Mutations in SDH subunits have been associated with tumorigenesis as well as mitochondrial diseases. In this project, we focused on the flavoprotein subunit A of SDH (SDHA) which is primarily associated with inherited mitochondrial disease [1] and investigated the consequences of this subunit loss in HEK cells (SDHA KO). <br> | |abstract='''Authors:''' [[Saucedo-Rodriguez Maria Jose]], [[Pecina Petr]], [[Cunatova Kristyna]], [[Vrbacky Marek]], [[Cajka T]], [[Mracek Tomas]], [[Pecinova Alena]]<br><br> | ||
Methods: We performed structural and functional characterizations of the SDHA KO model involving protein electrophoresis to study OXPHOS complexes and subcomplexes, label-free quantification of protein levels, measurement of cellular respiration using high-resolution respirometry and determination of NAD+/NADH levels.<br> | '''Introduction:''' Succinate dehydrogenase (SDH) connects the TCA cycle by oxidizing succinate to fumarate and the respiratory chain by transferring electrons to ubiquinone. Mutations in SDH subunits have been associated with tumorigenesis as well as mitochondrial diseases. In this project, we focused on the flavoprotein subunit A of SDH (SDHA) which is primarily associated with inherited mitochondrial disease [1] and investigated the consequences of this subunit loss in HEK cells (SDHA KO). <br> | ||
Results and discussion: Together with SDHA, other SDH subunits were downregulated as well, leading to the absence of assembled SDH. Moreover, a secondary downregulation of the majority of complex I and IV subunits was observed. The cellular respiratory capacity was severely decreased in the model, with SDH-dependent respiration completely abolished and complex I-dependent respiration attenuated reflecting the downregulation of respiratory chain complexes in general. Finally, the NAD+/NADH ratio was increased in SDHA KO compared to the controls, indicating complex rearrangement of the TCA. The SDHA KO cells thus represent a suitable model to study metabolic rewiring and the effect of pathogenic SDHA mutations.<br> | '''Methods:''' We performed structural and functional characterizations of the SDHA KO model involving protein electrophoresis to study OXPHOS complexes and subcomplexes, label-free quantification of protein levels, measurement of cellular respiration using high-resolution respirometry and determination of NAD<sup>+</sup>/NADH levels.<br> | ||
'''Results and discussion:''' Together with SDHA, other SDH subunits were downregulated as well, leading to the absence of assembled SDH. Moreover, a secondary downregulation of the majority of complex I and IV subunits was observed. The cellular respiratory capacity was severely decreased in the model, with SDH-dependent respiration completely abolished and complex I-dependent respiration attenuated reflecting the downregulation of respiratory chain complexes in general. Finally, the NAD<sup>+</sup>/NADH ratio was increased in SDHA KO compared to the controls, indicating complex rearrangement of the TCA. The SDHA KO cells thus represent a suitable model to study metabolic rewiring and the effect of pathogenic SDHA mutations.<br> | |||
<small> | <small> | ||
# Rustin, P., Munnich, A., & Rรถtig, A. (2002). Succinate dehydrogenase and human diseases: new insights into a well-known enzyme. https://doi.org/10.1038/sj.ejhg.5200793 ย | # Rustin, P., Munnich, A., & Rรถtig, A. (2002). Succinate dehydrogenase and human diseases: new insights into a well-known enzyme. https://doi.org/10.1038/sj.ejhg.5200793 ย | ||
</small> | </small> | ||
}} | }} | ||
=== Affiliations and acknowledgements === | |||
:::: Saucedo-Rodriguez Maria Jose<sup>1,2</sup>, Pecina P<sup>1</sup>, ฤunรกtovรก K<sup>1</sup>, Vrbackรฝ M<sup>1</sup>, ฤajka T<sup>3</sup>, Mrรกฤek T<sup>1</sup>, Pecinovรก A<sup>1</sup> | |||
::::# Laboratory of Bioenergetics, Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic | |||
::::# Faculty of Science, Charles University, Prague, Czech Republic | |||
::::# Laboratory of Translational Metabolism, Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic | |||
:::: Corresponding author: [email protected] | |||
:::: '''Funding:''' The project is supported by the Grant Agency of Charles University (GA UK No. 283423) | |||
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Revision as of 15:26, 17 April 2023
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Saucedo-Rodriguez 2023 MiP2023
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Link: MiP2023 Obergurgl AT
Saucedo-Rodriguez Maria Jose (2023)
Event: MiP2023 Obergurgl AT
Authors: Saucedo-Rodriguez Maria Jose, Pecina Petr, Cunatova Kristyna, Vrbacky Marek, Cajka T, Mracek Tomas, Pecinova Alena
Introduction: Succinate dehydrogenase (SDH) connects the TCA cycle by oxidizing succinate to fumarate and the respiratory chain by transferring electrons to ubiquinone. Mutations in SDH subunits have been associated with tumorigenesis as well as mitochondrial diseases. In this project, we focused on the flavoprotein subunit A of SDH (SDHA) which is primarily associated with inherited mitochondrial disease [1] and investigated the consequences of this subunit loss in HEK cells (SDHA KO).
Methods: We performed structural and functional characterizations of the SDHA KO model involving protein electrophoresis to study OXPHOS complexes and subcomplexes, label-free quantification of protein levels, measurement of cellular respiration using high-resolution respirometry and determination of NAD+/NADH levels.
Results and discussion: Together with SDHA, other SDH subunits were downregulated as well, leading to the absence of assembled SDH. Moreover, a secondary downregulation of the majority of complex I and IV subunits was observed. The cellular respiratory capacity was severely decreased in the model, with SDH-dependent respiration completely abolished and complex I-dependent respiration attenuated reflecting the downregulation of respiratory chain complexes in general. Finally, the NAD+/NADH ratio was increased in SDHA KO compared to the controls, indicating complex rearrangement of the TCA. The SDHA KO cells thus represent a suitable model to study metabolic rewiring and the effect of pathogenic SDHA mutations.
- Rustin, P., Munnich, A., & Rรถtig, A. (2002). Succinate dehydrogenase and human diseases: new insights into a well-known enzyme. https://doi.org/10.1038/sj.ejhg.5200793
Affiliations and acknowledgements
- Saucedo-Rodriguez Maria Jose1,2, Pecina P1, ฤunรกtovรก K1, Vrbackรฝ M1, ฤajka T3, Mrรกฤek T1, Pecinovรก A1
- Laboratory of Bioenergetics, Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic
- Faculty of Science, Charles University, Prague, Czech Republic
- Laboratory of Translational Metabolism, Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic
- Saucedo-Rodriguez Maria Jose1,2, Pecina P1, ฤunรกtovรก K1, Vrbackรฝ M1, ฤajka T3, Mrรกฤek T1, Pecinovรก A1
- Corresponding author: [email protected]
- Funding: The project is supported by the Grant Agency of Charles University (GA UK No. 283423)
Labels:
Event: E2
