Proteomic Analysis of Protein Expression and Oxidative Modification in R6/2 Transgenic Mice

Perluigi, Marzia; Poon, H. Fai; Maragos, William F.; Pierce, William M.; Klein, Jon B.; Calabrese, Vittorio; Cini, Chiara; Marco, C. De; Butterfield, D. Allan

doi:10.1074/mcp.m500090-mcp200

Cited by 168 publications

(156 citation statements)

References 75 publications

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“…Decreased PDH complex detected in HD patient's caudate nucleus was further correlated with increasing duration of the illness (Butterworth et al, 1985). Moreover, PDH expression was shown to decrease with age in the striatum of R6/2 transgenic mice (Perluigi et al, 2005). Previously, we also showed that HD cybrids present increased ROS production, compared to CTR cybrids (Ferreira et al, 2010); therefore we cannot exclude modifications in PDH activity resulting from enzyme oxidation.…”

Section: Discussionmentioning

confidence: 86%

Bioenergetic dysfunction in Huntington's disease human cybrids

Ferreira¹,

Cunha‐Oliveira²,

Nascimento³

et al. 2011

Experimental Neurology

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In this work we studied the mitochondrial-associated metabolic pathways in Huntington's disease (HD) versus control (CTR) cybrids, a cell model in which the contribution of mitochondrial defects from patients is isolated. HD cybrids exhibited an interesting increase in ATP levels, when compared to CTR cybrids. Concomitantly, we observed increased glycolytic rate in HD cybrids, as revealed by increased lactate/pyruvate ratio, which was reverted after inhibition of glycolysis. A decrease in glucose-6-phosphate dehydrogenase activity in HD cybrids further indicated decreased rate of the pentose-phosphate pathway. ATP levels of HD cybrids were significantly decreased under glycolysis inhibition, which was accompanied by a decrease in phosphocreatine. Nevertheless, pyruvate supplementation could not recover HD cybrids' ATP or phosphocreatine levels, suggesting a dysfunction in mitochondrial use of that substrate. Oligomycin also caused a decrease in ATP levels, suggesting a partial support of ATP generation by the mitochondria. Nevertheless, mitochondrial NADH/NAD t levels were decreased in HD cybrids, which was correlated with a decrease in pyruvate dehydrogenase activity and protein expression, suggesting decreased tricarboxylic acid cycle (TCA) input from glycolysis. Interestingly, the activity of alpha-ketoglutarate dehydrogenase, a critical enzyme complex that links the TCA to amino acid synthesis and degradation, was increased in HD cybrids. In accordance, mitochondrial levels of glutamate were increased and alanine was decreased, whereas aspartate and glutamine levels were unchanged in HD cybrids. Conversely, malate dehydrogenase activity from total cell extracts was unchanged in HD cybrids. Our results suggest that inherent dysfunction of mitochondria from HD patients affects cellular bioenergetics in an otherwise functional nuclear background.

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Section: Discussionmentioning

confidence: 86%

Bioenergetic dysfunction in Huntington's disease human cybrids

Ferreira¹,

Cunha‐Oliveira²,

Nascimento³

et al. 2011

Experimental Neurology

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“…and GST activity-We have previously shown that the oxidative modification of specific enzymes generally decreases their activity [82,92]. Therefore, in the present study we hypothesized that since increased oxidation leads to loss of enzymatic activity, then protection from oxidative damage could restore or maintain the activity of enzymes with up-regulated expression levels.…”

Section: Superoxide Dismutase (Sod)mentioning

confidence: 77%

“…Two-dimensional electrophoresis offers an excellent tool for the screening of abundant protein changes in various disease states [21,22,82,88]. In the present study, we investigated the pattern of protein expression in the parietal cortex in the four different groups.…”

Section: Protein Expression Levelsmentioning

confidence: 98%

“…We have also shown that α-enolase is oxidatively modified in AD and in various models of neurodegenerative disorders [32,82,83,90], indicating that this key protein is involved in several age-related neurodegenerative disorders. In addition, we have also shown that following caloric restriction in aging rats [93] and after treatment with lipoic acid in the SAMP8 mice [89], the specific carbonyl levels of α-enolase are significantly decreased leading us to believe that this protein may play a key role in the restoration of cognitive function.…”

Section: Energy Metabolismmentioning

confidence: 99%

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Proteomic identification of brain proteins in the canine model of human aging following a long-term treatment with antioxidants and a program of behavioral enrichment: Relevance to Alzheimer's disease

Opii

Joshi

Head

et al. 2008

Neurobiology of Aging

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176

135

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Aging and age-related disorders such as Alzheimer's disease (AD) are usually accompanied by oxidative stress as one of the main mechanisms contributing to neurodegeneration and cognitive decline. Aging canines develop cognitive dysfunction and neuropathology similar to those seen in humans, and the use of antioxidants results in reductions in oxidative damage and in improvement in cognitive function in this canine model of human aging. In the present study, the effect of a longterm treatment with an antioxidant fortified diet and a program of behavioral enrichment on oxidative damage was studied in aged canines. To identify the neurobiological mechanisms underlying these treatment effects, the parietal cortex from 23 beagle dogs (8.1-12.4 years) were treated for 2.8 years in one of four treatment groups: i.e., control food-control behavioral enrichment (CC); control food -behavioral enrichment (CE); antioxidant food-control behavioral enrichment (CA); and enriched environment -antioxidant fortified food (EA). We analyzed the levels of the oxidative stress biomarkers, i.e., protein carbonyls, 3-nitrotyroine (3NT), and the lipid peroxidation product, 4- Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Author ManuscriptNeurobiol Aging. Author manuscript; available in PMC 2009 January 1. Published in final edited form as:Neurobiol Aging. 2008 January ; 29(1): 51-70. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript hydroxynonenal (HNE), and observed a decrease in their levels on all treatments when compared to control, with the most significant effects found in the combined treatment, EA. Since EA treatment was most effective, we also carried out a comparative proteomics study to identify specific brain proteins that were differentially expressed and used a parallel redox proteomics approach to identify specific brain proteins that were less oxidized following EA. The specific protein carbonyl levels of glutamate dehydrogenase [NAD (P)], glyceraldehyde-3-phosphate dehydrogenase (GAPDH), α-enolase, neurofilament triplet L protein, glutathione S-transferase (GST) and fascin actin bundling protein were significantly reduced in brain of EA-treated dogs compared to control. We also observed significant increases in expression of Cu/Zn superoxide dismutase, fructose-bisphosphate aldolase C, creatine kinase, glutamate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase. The increased expression of these proteins and in particular Cu/Zn SOD correlated with improved cognitive function. In addition, there was a significant increas...

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“…Many proteins have been demonstrated to interact with WT Htt and/or mutant Htt, and their functions support some of the mechanisms perturbed in HD, which include transcription, signaling, trafficking/endocytosis, and metabolism/mitochondrial functions (5)(6)(7)(8)(9)(10). However, Htt's precise function has proven elusive, and our poor understanding of its function remains a limiting factor to the development of successful therapeutics (11).…”

mentioning

confidence: 99%

Huntington's disease protein contributes to RNA-mediated gene silencing through association with Argonaute and P bodies

Savas

Makusky

Ottosen

et al. 2008

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

155

113

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Huntington's disease is a dominant autosomal neurodegenerative disorder caused by an expansion of polyglutamines in the huntingtin (Htt) protein, whose cellular function remains controversial. To gain insight into Htt function, we purified epitope-tagged Htt and identified Argonaute as associated proteins. Colocalization studies demonstrated Htt and Ago2 to be present in P bodies, and depletion of Htt showed compromised RNA-mediated gene silencing. Mouse striatal cells expressing mutant Htt showed fewer P bodies and reduced reporter gene silencing activity compared with wild-type counterparts. These data suggest that the previously reported transcriptional deregulation in HD may be attributed in part to mutant Htt's role in post-transcriptional processes.microRNA ͉ poly-glutamine ͉ RNA interference ͉ post-transcriptional gene silencing ͉ neuronal RNA granule H untington's disease (HD) is caused by an expansion of the polymorphic stretch of uninterrupted CAG trinucleotide repeats in exon 1 of the IT15 gene, resulting in a long tract of polyglutamine (polyQ) in the N terminus of the HD protein huntingtin (Htt) (1). The human Htt protein contains 3,144 amino acids, and the polyQ stretch starts at the 18th amino acid, followed by a polyproline (polyP) sequence; the remaining portion of the protein is likely to be rich in ␣-helices, many of which compose HEAT repeats (2). The length of the glutamine repeats in the unaffected population varies from 6 to 35; HD is caused by expansion to 36 or more repeats (3, 4). HD is a member of a group of at least nine diseases caused by CAG repeat expansions that includes spinocerebellar ataxias (SCAs) 1-3, 6, 7, and 17, spinobulbar muscular atrophy, and dentatorubral-pallidoluysian atrophy. The protein that is subject to polyQ expansion in each disease appears to be unrelated, and despite their widespread expression in the brain and other tissues, mutation in each protein leads to distinct characteristic neurodegeneration patterns.Htt is expressed in a variety of tissues both within and outside of the nervous system. Many proteins have been demonstrated to interact with WT Htt and/or mutant Htt, and their functions support some of the mechanisms perturbed in HD, which include transcription, signaling, trafficking/endocytosis, and metabolism/mitochondrial functions (5-10). However, Htt's precise function has proven elusive, and our poor understanding of its function remains a limiting factor to the development of successful therapeutics (11). Proteolytic processing of WT and mutant Htt has been investigated extensively and appears to play a critical role in disease pathogenesis. Caspase-6 cleavage at amino acid 586 in mouse Htt and the release of the N-terminal Htt fragment are required for neuronal dysfunction and degeneration in an HD mouse model (12), but the basis for the need for this cleavage is unclear.We set out to search for binding proteins to elucidate the normal function of Htt. We chose a biochemical approach to purify proteins associated with WT and mutant Htt and ide...

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Proteomic Analysis of Protein Expression and Oxidative Modification in R6/2 Transgenic Mice

Cited by 168 publications

References 75 publications

Bioenergetic dysfunction in Huntington's disease human cybrids

Bioenergetic dysfunction in Huntington's disease human cybrids

Proteomic identification of brain proteins in the canine model of human aging following a long-term treatment with antioxidants and a program of behavioral enrichment: Relevance to Alzheimer's disease

Huntington's disease protein contributes to RNA-mediated gene silencing through association with Argonaute and P bodies

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