Characterizing Bcl-2 Family Protein Conformation and Oligomerization Using Cross-Linking and Antibody Gel-Shift in Conjunction with Native PAGE

Dewson, Grant

doi:10.1007/978-1-4939-3581-9_14

Cited by 5 publications

(2 citation statements)

References 22 publications

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“…The Bcl-2 family consists of three subgroups, pro-survival proteins (BCL2, BCLxl [BCL2L1], BCLw [BCL2L2], MCL-1, and A1), multi-domain pro-apoptotic proteins (BAX and BAK), and BH3 domain-only pro-apoptotic proteins (BIM, PUMA [BBC3], BID, BAD, BIK, BMF, HRK, and NOXA [PMAIP1]) ( 17 – 19 ). As an important anti-apoptotic protein in the Bcl-2 family, inhibition of MCL-1 promotes cell death through the mitochondrial pathway and there is some data suggesting it is also involved in the apoptosis that occurs during cerebral ischemic reperfusion ( 20 ). Based on these data and our results, we hypothesize that the interaction between miR-29b and MCL-1 is a critical factor during neural cell apoptosis.…”

Section: Discussionmentioning

confidence: 99%

miR‑29b affects neurocyte apoptosis by targeting MCL‑1 during cerebral ischemia/reperfusion injury

Huang

Lu²,

Jiang

et al. 2018

Exp Ther Med

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The present study aimed to determine whether an miRNA (miR)-29b inhibitor protected against cerebral ischemia/reperfusion (I/R) injury in vitro and to investigate the underlying mechanisms. As a model for induced cerebral IR injury, N2a cells were exposed to an oxygen-glucose deprivation/reoxygenation (OGD/R) environment. Using this model, it was demonstrated that miR-29b was significantly upregulated compared with cells in a normal environment. The interactions between miR-29b and myeloid cell leukemia sequence (MCL)-1 were then investigated using dual-luciferase assays, revealing a strong regulation of MCL-1 through the 3′untranslated region. Using the OGD/R model, the present study additionally examined the effects of miR-29b and miR-29b inhibitor on cell viability and apoptosis using Cell Counting kit 8 and flow cytometry assays, respectively. miR-29b transfection led to increased N2a cell apoptosis and reduced cell viability under an OGD/R environment. However, this effect was reversed by the miR-29b inhibitor. Finally, the effects of miR-29b on the expression of several Wnt-associating proteins were examined. It was observed that B cell lymphoma-2 was inhibited by miR-29b, as was MCL-1, whereas caspase-3 expression was promoted. The miR-29b inhibitor demonstrated the opposite effect. Overall, miR-29b promoted neurocyte apoptosis by targeting MCL-1 during cerebral I/R injury. The results of the present study suggest a potential novel therapeutic target for the treatment of ischemic stroke.

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Section: Discussionmentioning

confidence: 99%

miR‑29b affects neurocyte apoptosis by targeting MCL‑1 during cerebral ischemia/reperfusion injury

Huang

Lu²,

Jiang

et al. 2018

Exp Ther Med

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“…In native PAGE, denaturing agents such as sodium dodecyl sulphate (SDS) are omitted. As such, the proteins are folded, and protein complexes will exhibit electrophoretic shifts (generally reduced mobility) compared to negative controls (Dewson, 2016;Krause, 2006).…”

Section: Afcsos2 Interacts With Both Rubisco and Afcsos1amentioning

confidence: 99%

Towards the in vitro reconstitution of proteobacterial carboxysomes

How¹

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Methods to Probe Conformational Activation and Mitochondrial Activity of Proapoptotic BAK

Singh

Moldoveanu

2018

Methods in Molecular Biology

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Characterizing Bcl-2 Family Protein Conformation and Oligomerization Using Cross-Linking and Antibody Gel-Shift in Conjunction with Native PAGE

Cited by 5 publications

References 22 publications

miR‑29b affects neurocyte apoptosis by targeting MCL‑1 during cerebral ischemia/reperfusion injury

miR‑29b affects neurocyte apoptosis by targeting MCL‑1 during cerebral ischemia/reperfusion injury

Towards the in vitro reconstitution of proteobacterial carboxysomes

Methods to Probe Conformational Activation and Mitochondrial Activity of Proapoptotic BAK

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