Interactions of 17β-Hydroxysteroid Dehydrogenase Type 10 and Cyclophilin D in Alzheimer's Disease

Krištofiková, Zdeňka; Špringer, Tomáš; Gedeonova, Erika; Hofmannova, Adéla; Říčný, Jan; Hromádková, Lenka; Vyhnálek, Martin; Laczó, Jan; Nikolai, Tomáš; Hort, Jakub; Petrásek, Tomáš; Stuchlı́k, Aleš; Valeš, Karel; Klaschka, Jan; Homola, Jiřı́

doi:10.1007/s11064-020-02970-y

Cited by 8 publications

(6 citation statements)

References 42 publications

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“…As the absolute concentrations of Aβ, cypD and 17β-HSD10 in mitochondria are still a subject of research, in our simulations, we assumed a ratio of cypD:17β-HSD10 of 1:7 [ 43 ] and much smaller initial concentrations of Aβ in comparison to cypD and 17β-HSD10. The particular parameters used in the model are summarized in Table S1 in Supplementary information .…”

Section: Resultsmentioning

confidence: 99%

Study of Biomolecular Interactions of Mitochondrial Proteins Related to Alzheimer’s Disease: Toward Multi-Interaction Biomolecular Processes

et al. 2020

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Progressive mitochondrial dysfunction due to the accumulation of amyloid beta (Aβ) peptide within the mitochondrial matrix represents one of the key characteristics of Alzheimer’s disease (AD) and appears already in its early stages. Inside the mitochondria, Aβ interacts with a number of biomolecules, including cyclophilin D (cypD) and 17β-hydroxysteroid dehydrogenase type 10 (17β-HSD10), and affects their physiological functions. However, despite intensive ongoing research, the exact mechanisms through which Aβ impairs mitochondrial functions remain to be explained. In this work, we studied the interactions of Aβ with cypD and 17β-HSD10 in vitro using the surface plasmon resonance (SPR) method and determined the kinetic parameters (association and dissociation rates) of these interactions. This is the first work which determines all these parameters under the same conditions, thus, enabling direct comparison of relative affinities of Aβ to its mitochondrial binding partners. Moreover, we used the determined characteristics of the individual interactions to simulate the concurrent interactions of Aβ with cypD and 17β-HSD10 in different model situations associated with the progression of AD. This study not only advances the understanding of Aβ-induced processes in mitochondria during AD, but it also provides a new perspective on research into complex multi-interaction biomolecular processes in general.

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Section: Resultsmentioning

confidence: 99%

Study of Biomolecular Interactions of Mitochondrial Proteins Related to Alzheimer’s Disease: Toward Multi-Interaction Biomolecular Processes

et al. 2020

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“…Samples were prepared according to the general procedure as described by ( Kristofikova et al, 2020 ) but with immediate processing without storage. The resin was TAAB 813 (TAAB, Aldermaston) and after sectioning samples were stained with EM Stain 336 (Agar Scientific Ltd., Stansted) and Reynold’s stain.…”

Section: Methodsmentioning

confidence: 99%

Rapid isolation of respiring skeletal muscle mitochondria using nitrogen cavitation

Younis

Lavery²,

Christian³

et al. 2023

Front. Physiol.

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Methods of isolating mitochondria commonly utilise mechanical force and shear stress to homogenize tissue followed by purification by multiple rounds of ultracentrifugation. Existing protocols can be time-consuming with some physically impairing integrity of the sensitive mitochondrial double membrane. Here, we describe a method for the recovery of intact, respiring mitochondria from murine skeletal muscle tissue and cell lines using nitrogen cavitation. This protocol results in high-yield, pure and respiring mitochondria without the need for purification gradients or ultracentrifugation. The protocol takes under an hour and requires limited specialised equipment. Our methodology is successful in extracting mitochondria of both cell extracts and skeletal muscle tissue. This represents an improved yield in comparison to many of the existing methods. Western blotting and electron microscopy demonstrate the enrichment of mitochondria with their ultrastructure well-preserved and an absence of contamination from cytoplasmic or nuclear fractions. Using respirometry analysis we show that mitochondria extracted from murine skeletal muscle cell lines (C2C12) and tibialis anterior tissue have an appropriate respiratory control ratio. These measures are indicative of healthy coupled mitochondria. Our method successfully demonstrates the rapid isolation of functional mitochondria and will benefit researchers studying mitochondrial bioenergetics as well as providing greater throughput and application for time-sensitive assays.

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“…Samples were prepared according to the general procedure as described by (12) but with immediate processing without storage. The resin was TAAB 813 (TAAB, Aldermaston) and after sectioning samples were stained with EM Stain 336 (Agar Scientific Ltd., Stansted) and Reynold’s stain.…”

Section: Methodsmentioning

confidence: 99%

Rapid isolation of respiring skeletal muscle mitochondria using nitrogen cavitation

Younis

Lavery

Christian

et al. 2022

Preprint

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Background Methods of isolating mitochondria commonly utilize mechanical force and shear stress to homogenize tissue followed by purification by multiple rounds of ultracentrifugation. Existing protocols can be time-consuming with some physically impairing integrity of the sensitive mitochondrial double membrane. Methods Here, we describe a method for the recovery of intact, respiring mitochondria from murine skeletal muscle tissue and cell lines using nitrogen cavitation in combination with differential centrifugation. Results This protocol results in high yield, pure and respiring mitochondria without the need for purification gradients or ultracentrifugation. The protocol takes under an hour and requires limited specialised equipment. Our methodology is successful in extracting mitochondria of both cell extracts and skeletal muscle tissue. This represents an improved yield in comparison to many of the existing methods. Western blotting and electron microscopy demonstrate an enrichment of mitochondria with their ultrastructure well-preserved and an absence of contamination from cytoplasmic or nuclear fractions. Using respirometry analysis we show that mitochondria extracted from the murine skeletal muscle cell lines and tibialis anterior have an appropriate respiratory control ratio. These measures are indicative of healthy coupled mitochondria. Conclusion Our method successfully demonstrates the rapid isolation of functional mitochondria and will benefit researchers studying mitochondrial bioenergetics as well as providing greater throughput and application for time-sensitive assays.

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Interactions of 17β-Hydroxysteroid Dehydrogenase Type 10 and Cyclophilin D in Alzheimer's Disease

Cited by 8 publications

References 42 publications

Study of Biomolecular Interactions of Mitochondrial Proteins Related to Alzheimer’s Disease: Toward Multi-Interaction Biomolecular Processes

Study of Biomolecular Interactions of Mitochondrial Proteins Related to Alzheimer’s Disease: Toward Multi-Interaction Biomolecular Processes

Rapid isolation of respiring skeletal muscle mitochondria using nitrogen cavitation

Rapid isolation of respiring skeletal muscle mitochondria using nitrogen cavitation

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