Structure and function of mitochondrial membrane protein complexes

Kühlbrandt, Werner

doi:10.1186/s12915-015-0201-x

Cited by 530 publications

(430 citation statements)

References 64 publications

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“…The mitochondrial respiratory complexes catalyze the transfer of electrons from NADH to create a proton gradient across the inner membrane and drive ATP production via ATP synthases (33,34). A serial assay of mitochondrial function was conducted, and mitochondrial damages were confirmed in both TG02 and TMZ treatment in our studies (Fig.…”

Section: Discussionmentioning

confidence: 58%

Novel Targeting of Transcription and Metabolism in Glioblastoma

Chen

Wang

et al. 2018

Clinical Cancer Research

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Purpose: Glioblastoma (GBM) is highly resistant to treatment, largely due to disease heterogeneity and resistance mechanisms. We sought to investigate a promising drug that can inhibit multiple aspects of cancer cell survival mechanisms and become an effective therapeutic for GBM patients.Experimental Design: To investigate TG02, an agent with known penetration of the blood-brain barrier, we examined the effects as single agent and in combination with temozolomide, a commonly used chemotherapy in GBM. We used human GBM cells and a syngeneic mouse orthotopic GBM model, evaluating survival and the pharmacodynamics of TG02. Mechanistic studies included TG02-induced transcriptional regulation, apoptosis, and RNA sequencing in treated GBM cells as well as the investigation of mitochondrial and glycolytic function assays.Results: We demonstrated that TG02 inhibited cell proliferation, induced cell death, and synergized with temozolomide in GBM cells with different genetic background but not in astrocytes. TG02-induced cytotoxicity was blocked by the overexpression of phosphorylated CDK9, suggesting a CDK9-dependent cell killing. TG02 suppressed transcriptional progression of antiapoptotic proteins and induced apoptosis in GBM cells. We further demonstrated that TG02 caused mitochondrial dysfunction and glycolytic suppression and ultimately ATP depletion in GBM. A prolonged survival was observed in GBM mice receiving combined treatment of TG02 and temozolomide. The TG02-induced decrease of CDK9 phosphorylation was confirmed in the brain tumor tissue.Conclusions: TG02 inhibits multiple survival mechanisms and synergistically decreases energy production with temozolomide, representing a promising therapeutic strategy in GBM, currently under investigation in an ongoing clinical trial.

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Section: Discussionmentioning

confidence: 58%

Novel Targeting of Transcription and Metabolism in Glioblastoma

Chen

Wang

et al. 2018

Clinical Cancer Research

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“…S1D), that identifies an interfacial region for these proteins near the N terminus (K119) of MIC60. As a final point, we observed interactions between the MICOS/MIB complex protein MIC27 and subunits ATPA, ATP5F1, and ATP5J of the F 1 domain of ATP synthase, including two ATP synthase stator domain proteins, ATP5F1 and ATP5J (50,51). These ATP synthase interactions with MICOS/MIB provide structural insight (stator binding) consistent with the reported roles of MICOS/MIB complexes in coordinating with OXPHOS complexes, including ATP synthase, to define the architecture of the inner mitochondrial membrane.…”

Section: Protein Sitementioning

confidence: 91%

Mitochondrial protein interactome elucidated by chemical cross-linking mass spectrometry

Schweppe

Chavez

Lee

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

174

206

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Mitochondrial protein interactions and complexes facilitate mitochondrial function. These complexes range from simple dimers to the respirasome supercomplex consisting of oxidative phosphorylation complexes I, III, and IV. To improve understanding of mitochondrial function, we used chemical cross-linking mass spectrometry to identify 2,427 cross-linked peptide pairs from 327 mitochondrial proteins in whole, respiring murine mitochondria. In situ interactions were observed in proteins throughout the electron transport chain membrane complexes, ATP synthase, and the mitochondrial contact site and cristae organizing system (MICOS) complex. Cross-linked sites showed excellent agreement with empirical protein structures and delivered complementary constraints for in silico protein docking. These data established direct physical evidence of the assembly of the complex I-III respirasome and enabled prediction of in situ interfacial regions of the complexes. Finally, we established a database and tools to harness the cross-linked interactions we observed as molecular probes, allowing quantification of conformation-dependent protein interfaces and dynamic protein complex assembly. mitochondria | mass spectrometry | interactome | cross-linking | protein interaction reporter M itochondrial proteins play a diverse role in cellular biology and disease. Mitochondrial dysfunction directly causes multiple inherited diseases (1) and is implicated in common diseases, including neurological developmental disorders (2, 3), neurodegenerative and cardiovascular diseases (4-6), diabetes (7), asthma (8), cancer (9), and age-related disease (10). In mammals, these organelles have evolved to retain more than 1,000 proteins that interact within a complex, i.e., dual membrane architecture (11,12). Within the mitochondrial proteome, the "powerhouse" functions are carried out by the core constituents of the oxidative phosphorylation (OXPHOS) system [complexes I-IV of the electron transport chain (ETC) and ATP synthase (complex V)]. These proteins are necessary for creation of the mitochondrial electrochemical gradient that powers synthesis of ATP. This system includes critical protein-protein interactions within individual OXPHOS complexes as well as "supercomplex" interactions between ETC complexes I, III, and IV in the respirasome (13). Deficient supercomplex formation has been proposed as a critical mitochondrial defect in failing hearts (5,6,14,15), and dynamic rearrangement of supercomplexes has been implicated in noncanonical mitochondrial functions such as antibacterial innate immune responses (16). Assessing these interactions is further complicated by regulatory posttranslational modification and conformational changes of mitochondrial proteins (17)(18)(19)(20). Advances in this area have been impeded, in part, by the lack of large-scale detection of dynamic, sometimes transient, interactions between membrane proteins. Thus, large-scale determination of the protein interactome within mitochondria would provide a valuable tool to adv...

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“…Mitochondria are the main sites of ATP generation (Kuhlbrandt, 2015). High levels of UA were previously found to induce mitochondrial dysfunction, which may lead to excess ROS generation.…”

Section: Discussionmentioning

confidence: 99%

“…Aberrant UA metabolism can cause hepatocyte injury by inducing oxidative stress. The mitochondria is the primary organelle involved in restoring oxygen molecules, and is an important source of reactive oxygen species (ROS) (Kuhlbrandt, 2015). Purine metabolism occurs predominantly in hepatocytes, which contain 500-1000 mitochondria; therefore, these cells play a major role in oxidative stress.…”

Section: Introductionmentioning

confidence: 99%

Effect of uric acid on mitochondrial function and oxidative stress in hepatocytes

et al. 2016

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ABSTRACT. Here, we investigated the effect of uric acid (UA) on hepatocyte mitochondria. Hepatocytes cultured in vitro were treated with varying concentrations of UA. The change in apoptotic activity was detected by flow cytometry. The DNA damage index 8-hydroxy-deoxyguanosine (8-OHdG) and mitochondrial function indices succinate dehydrogenase (SDH), cytochrome C oxidase (CCO), and adenosine triphosphate (ATP) were detected by enzyme assays. Reactive oxygen species (ROS) accumulation was confirmed by a dichloro-dihydrofluorescein diacetate assay. We observed an increase in apoptotic activity, ROS accumulation, and 8-OHdG activity in hepatocytes treated with UA for extended periods, indicating DNA damage; specifically, we observed a significant increase in these activities 48, 72, and 96 h after UA addition, compared to those observed at 24 h (P < 0.05). Cells treated with 30 mg/dL UA for 96 h showed a peak in apoptotic activity. We also observed a significant decrease in ATP, SDH, and CCO activities with the increase in uric acid concentration over time. Cells treated with 30 mg/dL UA for 96 h showed the highest ATP levels, while SDH and CCO activities at 48, 72, and 96 h post-UA treatment were significantly lower than those at 24 h (P < 0.01). Moreover, cells treated with 30 mg/dL UA showed a 0.02 ± 0.02 and 0.15 ± 0.01 mmol/ mg/min decrease in SDH and CCO levels after 72 h. Therefore, we concluded that high concentrations of UA may induce oxidative stress in hepatocyte mitochondria, increasing ROS production and ultimately resulting in mitochondrial damage.

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Structure and function of mitochondrial membrane protein complexes

Cited by 530 publications

References 64 publications

Novel Targeting of Transcription and Metabolism in Glioblastoma

Novel Targeting of Transcription and Metabolism in Glioblastoma

Mitochondrial protein interactome elucidated by chemical cross-linking mass spectrometry

Effect of uric acid on mitochondrial function and oxidative stress in hepatocytes

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