Synthesis of Ventricular Mitochondrial Proteins In Vivo: Effect of Acute Ethanol Toxicity

Mitochondria are recognized as signaling organelles because, under stress, mitochondria can trigger various signaling pathways to coordinate the cell's response. The specific pathway(s) engaged by mitochondria in response to mitochondrial energy defects in vivo and in high-energy tissues like the heart are not fully understood. Here, we investigated cardiac pathways activated in response to mitochondrial energy dysfunction by studying mice with cardiomyocyte-specific loss of the mitochondrial phosphate carrier (SLC25A3), an established model that develops cardiomyopathy as a result of defective mitochondrial ATP synthesis. Mitochondrial energy dysfunction induced a striking pattern of acylome remodeling, with significantly increased post-translational acetylation and malonylation. Mass spectrometry-based proteomics further revealed that energy dysfunction-induced remodeling of the acetylome and malonylome preferentially impacts mitochondrial proteins. Acetylation and malonylation modified a highly interconnected interactome of mitochondrial proteins, and both modifications were present on the enzyme isocitrate dehydrogenase 2 (IDH2). Intriguingly, IDH2 activity was enhanced in SLC25A3-deleted mitochondria, and further study of IDH2 sites targeted by both acetylation and malonylation revealed that these modifications can have site-specific and distinct functional effects. Finally, we uncovered a novel crosstalk between the two modifications, whereby mitochondrial energy dysfunction-induced acetylation of sirtuin 5 (SIRT5), inhibited its function. Because SIRT5 is a mitochondrial deacylase with demalonylase activity, this finding suggests that acetylation can modulate the malonylome. Together, our results position acylations as an arm of the mitochondrial response to energy dysfunction and suggest a mechanism by which focal disruption to the energy production machinery can have an expanded impact on global mitochondrial function.

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Differences in IL-10 and IL-12 production patterns and differences in the effects of acute ethanol treatment on macrophages in vivo and in vitro

Alcohol ◽

10.1016/j.alcohol.2005.09.004 ◽

2005 ◽

Vol 37 (1) ◽

pp. 1-8 ◽

Cited By ~ 17

Author(s):

Stephen B. Pruett ◽

Ruping Fan ◽

Qiang Zheng ◽

Carlton Schwab

Keyword(s):

Ethanol Treatment ◽

Acute Ethanol

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In vivo voltammetric evidence that locus coeruleus activation predominantly releases norepinephrine in the infralimbic cortex: Effect of acute ethanol

Synapse ◽

10.1002/syn.22080 ◽

2018 ◽

Vol 73 (4) ◽

pp. e22080 ◽

Cited By ~ 3

Author(s):

Alex L. Deal ◽

Maria A. Mikhailova ◽

Valentina P. Grinevich ◽

Jeff L. Weiner ◽

Raul R. Gainetdinov ◽

...

Keyword(s):

Locus Coeruleus ◽

Infralimbic Cortex ◽

Acute Ethanol

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Acute Ethanol Administration Rapidly Increases Phosphorylation of Conventional Protein Kinase C in Specific Mammalian Brain Regions in Vivo

Alcoholism Clinical and Experimental Research ◽

10.1111/j.1530-0277.2007.00423.x ◽

2007 ◽

Vol 31 (7) ◽

pp. 1259-1267 ◽

Cited By ~ 22

Author(s):

Mary Beth Wilkie ◽

Joyce Besheer ◽

Stephen P. Kelley ◽

Sandeep Kumar ◽

Todd K. O'Buckley ◽

...

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Brain Regions ◽

Ethanol Administration ◽

Mammalian Brain ◽

Kinase C ◽

Acute Ethanol

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Effects of acute ethanol on ventricular myofibrillary protein synthesis in vivo in normotensive and hypertensive rats

Addiction Biology ◽

10.1080/13556219772895 ◽

1997 ◽

Vol 2 (1) ◽

pp. 87-93 ◽

Cited By ~ 10

Author(s):

TAHIR SIDDIQ ◽

GURJINDER SANDHU ◽

PETER J. RICHARDSON ◽

VICTOR R. PREEDY

Keyword(s):

Protein Synthesis ◽

Hypertensive Rats ◽

Acute Ethanol

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Acute Ethanol Gavage Attenuates Hemorrhage/Resuscitation-Induced Hepatic Oxidative Stress in Rats

Oxidative Medicine and Cellular Longevity ◽

10.1155/2012/983427 ◽

2012 ◽

Vol 2012 ◽

pp. 1-10 ◽

Cited By ~ 10

Author(s):

B. Relja ◽

K. Wilhelm ◽

M. Wang ◽

D. Henrich ◽

I. Marzi ◽

...

Keyword(s):

Oxidative Stress ◽

Ethanol Intoxication ◽

Arterial Blood ◽

Inos Protein Expression ◽

Acute Ethanol ◽

Acute Fatty Liver ◽

Acute Ethanol Intoxication ◽

Subsequent Resuscitation

Acute ethanol intoxication increases the production of reactive oxygen species (ROS). Hemorrhagic shock with subsequent resuscitation (H/R) also induces ROS resulting in cellular and hepatic damagein vivo. We examined the role of acute ethanol intoxication upon oxidative stress and subsequent hepatic cell death after H/R. 14 h before H/R, rats were gavaged with single dose of ethanol or saline (5 g/kg, EtOH and ctrl; H/R_EtOH or H/R_ctrl, resp.). Then, rats were hemorrhaged to a mean arterial blood pressure of30±2 mmHg for 60 min and resuscitated. Two control groups underwent surgical procedures without H/R (sham_ctrl and sham_EtOH, resp.). Liver tissues were harvested at 2, 24, and 72 h after resuscitation. EtOH-gavage induced histological picture of acute fatty liver. Hepatic oxidative (4-hydroxynonenal, 4-HNE) and nitrosative (3-nitrotyrosine, 3-NT) stress were significantly reduced in EtOH-gavaged rats compared to controls after H/R. Proapoptotic caspase-8 and Bax expressions were markedly diminished in EtOH-gavaged animals compared with controls 2 h after resuscitation. EtOH-gavage increased antiapoptotic Bcl-2 gene expression compared with controls 2 h after resuscitation. iNOS protein expression increased following H/R but was attenuated in EtOH-gavaged animals after H/R. Taken together, the data suggest that acute EtOH-gavage may attenuate H/R-induced oxidative stress thereby reducing cellular injury in rat liver.

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The synthesis of murine ferrochelatase in vitro and in vivo

Biochemical Journal ◽

10.1042/bj2540799 ◽

1988 ◽

Vol 254 (3) ◽

pp. 799-803 ◽

Cited By ~ 29

Author(s):

S R Karr ◽

H A Dailey

Keyword(s):

Membrane Protein ◽

Biosynthetic Pathway ◽

Integral Membrane Protein ◽

Mitochondrial Proteins ◽

Isolated Mitochondria ◽

Mitochondrial Inner Membrane ◽

Carbonyl Cyanide ◽

A Cell

Ferrochelatase (protohaem ferro-lyase, EC 4.99.1.1), the terminal enzyme of the haem-biosynthetic pathway, is an integral membrane protein of the mitochondrial inner membrane. When murine erythroleukaemia cells are labelled in vivo with [35S]methionine, lysed, and the extract is immunoprecipitated with rabbit anti-(mouse ferrochelatase) antibody, a protein of Mr 40,000 is isolated. However, when isolated mouse RNA is translated in a cell-free reticulocyte extract, a protein of Mr 43,000 is isolated. Incubation of this Mr 43,000 protein with isolated mitochondria resulted in processing of the Mr 43,000 precursor to the Mr 40,000 mature-sized protein. Addition of carbonyl cyanide m-chlorophenylhydrazone and/or phenanthroline inhibits this processing. These data indicate that ferrochelatase, like most mitochondrial proteins, is synthesized in the cytoplasm as a larger precursor and is then translocated and processed to a mature-sized protein in an energy-required step.

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High fat diet induces dysregulation of hepatic oxygen gradients and mitochondrial function in vivo

Biochemical Journal ◽

10.1042/bj20080868 ◽

2008 ◽

Vol 417 (1) ◽

pp. 183-193 ◽

Cited By ~ 174

Author(s):

Sudheer K. Mantena ◽

Denty Paul Vaughn ◽

Kelly K. Andringa ◽

Heather B. Eccleston ◽

Adrienne L. King ◽

...

Keyword(s):

Nitric Oxide ◽

High Fat Diet ◽

Protein Modification ◽

Control Diet ◽

Critical Role ◽

Liver Mitochondria ◽

Mitochondrial Proteins ◽

High Fat ◽

Alcoholic Fatty Liver

NAFLD (non-alcoholic fatty liver disease), associated with obesity and the cardiometabolic syndrome, is an important medical problem affecting up to 20% of western populations. Evidence indicates that mitochondrial dysfunction plays a critical role in NAFLD initiation and progression to the more serious condition of NASH (non-alcoholic steatohepatitis). Herein we hypothesize that mitochondrial defects induced by exposure to a HFD (high fat diet) contribute to a hypoxic state in liver and this is associated with increased protein modification by RNS (reactive nitrogen species). To test this concept, C57BL/6 mice were pair-fed a control diet and HFD containing 35% and 71% total calories (1 cal≈4.184 J) from fat respectively, for 8 or 16 weeks and liver hypoxia, mitochondrial bioenergetics, NO (nitric oxide)-dependent control of respiration, and 3-NT (3-nitrotyrosine), a marker of protein modification by RNS, were examined. Feeding a HFD for 16 weeks induced NASH-like pathology accompanied by elevated triacylglycerols, increased CYP2E1 (cytochrome P450 2E1) and iNOS (inducible nitric oxide synthase) protein, and significantly enhanced hypoxia in the pericentral region of the liver. Mitochondria from the HFD group showed increased sensitivity to NO-dependent inhibition of respiration compared with controls. In addition, accumulation of 3-NT paralleled the hypoxia gradient in vivo and 3-NT levels were increased in mitochondrial proteins. Liver mitochondria from mice fed the HFD for 16 weeks exhibited depressed state 3 respiration, uncoupled respiration, cytochrome c oxidase activity, and mitochondrial membrane potential. These findings indicate that chronic exposure to a HFD negatively affects the bioenergetics of liver mitochondria and this probably contributes to hypoxic stress and deleterious NO-dependent modification of mitochondrial proteins.

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Chronic and acute ethanol impair the in vivo glucose uptake by lactating rat mammary gland

Bioscience Reports ◽

10.1007/bf01116750 ◽

1987 ◽

Vol 7 (10) ◽

pp. 777-781 ◽

Cited By ~ 1

Author(s):

S. Vilaró ◽

O. Viñas ◽

E. Herrera ◽

X. Remesar

Keyword(s):

Mammary Gland ◽

Glucose Uptake ◽

Ketone Bodies ◽

Direct Effects ◽

Acute Ethanol ◽

Rat Mammary Gland

Chronic and acute ethanol treatments increased the 3-hydroxybutyrate uptake by lactating rat mammary gland as a consequence of its high afferent concentration, without changing its relative extraction. The uptake of glucose was inhibited in the ethanol treated animals due to intrinsic alterations in the mammary gland metabolism as indicated by the decreased relative extraction and unchanged afferent concentration. These results would suggest that the elevated uptake of ketone bodies in ethanol-treated rats can be responsible, at least in part, for the decrease in glucose uptake by lactating rat mammary gland, although other direct effects of ethanol may be implied.

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