Biochemical studies have shown that microsomes represent an important subcellular fraction for determining 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) effects. Proteomic analysis by two-dimensional gel-mass spectrometry of liver microsomes was undertaken to gain new insight into the actions of TCDD in male and female rats. Proteomic analysis showed TCDD induced several xenobiotic metabolism enzymes as well as a protein at 90kDa identified by mass spectrometry as IkappaB kinase beta/IKK2. This observation led to the discovery of other NF-kappaB binding proteins and kinases in microsomes and effects by TCDD. Western blotting for IKK and IkappaB family members in microsomes showed a distinct pattern from cytosol. IKK1 and IKK2 were both present in microsomes and were catalytically active although, unlike cytosol, IKKgamma/NEMO was not detectable. TCDD exposure produced an elevation in cytosolic and microsomal IKK activity of both genders. The NF-kappaB binding proteins IkappaBbeta and IkappaBgamma were prevalent in microsomes, while IkappaBalpha and IkappaB epsilon proteins were absent. TCDD treatment produced hyperphosphorylation of microsomal IkappaBbeta in both sexes with females being most sensitive. In cytosol, IkappaBalpha, IkappaBbeta, and IkappaB epsilon, but not IkappaBgamma, were clearly observed but were not changed by TCDD. Overall, proteomic analysis indicated the presence of NF-kappaB pathway members in microsomes, selectively altered by dioxin, which may influence immune and inflammatory responses within the liver.
"). Prefractionation of the original sample is an appropriate means to arrive at subproteomes, for example, of subcellular organelles (Bruno et al., 2002; Lasserre et al., 2012). In addition, chromatographic steps (1) based on different separation criteria and (2) taking advantage of specific protein properties, such as isoelectric point, hydrophobicity , and interaction with affinity matrices, may reduce sample complexity and reveal otherwise hidden proteins (Miller, 2011; Capriotti et al., 2012). "
[Show abstract][Hide abstract] ABSTRACT: Proteomics has the potential to elucidate complex patterns of toxic action attributed to its unique holistic a posteriori approach. In the case of toxic compounds for which the mechanism of action is not completely understood, a proteomic approach may provide valuable mechanistic insight. This review provides an overview of currently available proteomic techniques, including examples of their application in toxicological in vivo and in vitro studies. Future perspectives for a wider application of state-of-the-art proteomic techniques in the field of toxicology are discussed. The examples concern experiments with dioxins, polychlorinated biphenyls, and polybrominated diphenyl ethers as model compounds, as they exhibit a plethora of sublethal effects, of which some mechanisms were revealed via successful proteomic studies. Generally, this review shows the added value of including proteomics in a modern tool box for toxicological studies.
Journal of Toxicology and Environmental Health Part B 05/2014; 17(4):225-46. DOI:10.1080/10937404.2014.904730 · 4.97 Impact Factor
"Rat kidney; plasma ↑ Plasma kininogen in renal toxicity; plasma biomarkers (Bruno et al., 2002) 2D-MS TCDD Rat liver ER NFkB proteins, IKBs, IKKs found in ER (Castegna et al., 2003) 2D-MS ONOO- Human brain Six targets of protein nitration IDed in Alzheimer brains (Charlwood et al., 2002) 2D-MS Gentamicin Rat kidney 20 proteins IDed; mitochondria dysfunction in renal cortex (Cutler et al., 1999) 2D-MS Puromycin Rat urine Proteomics/metabolomics to study renal glomerular toxicity (Dapas et al., 2003) 2D-MS GT oligomers Human lympocytes Basic isoform of eEF1A IDed in GT oligomer cytotoxicity-complex (Dare et al., 2002) SELDI TMPD Rat urine Parvalbumin-α biomarker of skeletal muscle toxicity (Drake et al., 2003) 2D-MS JP-8 jet fuel Mouse lung Proteins IDed during apoptosis and edema after exposure (Fennell et al., 2003) LC-MS/MS Acrylamides Rat blood Hgb adducted sites IDed (Fessler et al., 2002) 2D-MS LPS; Human neutro-phils, in vitro DNA array/proteomics; ↑ in inflammation signaling, cytoskeletal proteins IDed (Fountoulakis et al., 2000) 2D-MS APAP, AMAP Mouse liver 35 proteins IDed; altered proteins are known targets for adducts (Gao et al., 2004) MudPIT 20 cytotoxic, nontoxic isomers Human hepatocytes ↑ BMS-PTX-265 and BMS-PTX-837 proteins released in culture medium relate to toxicity (Heijne et al., 2003) 2D-MS Bromobenzene Rat liver DNA array/proteomics; IDed proteins infer degradation, oxidative stress from toxicity (Hestvik et al., 2003) Anti-Phos Ab array BCG Mycobact infection of host Monocyte THP-1 cells Activation of SAP kinase cJun, ↓PKC varε; ↑ α-adducin, GSK-3β by BCG infection (Hirata et al., 2003) Ab array cytokine Cardiotoxin Mouse skeletal muscle DNA array/proteomics; ↑ of osteopontin, C10/CCL6 with muscle injury (Hogstrand et al., 2002) SELDI Zinc Trout, gill DNA array/proteomics; proteins altered by SELDI; no protein ID (Iida et al., 2003) 2D-MS Oxazepam, Wyeth 14643 Mouse liver DNA array/proteomics; subcellular fractions, protein IDs unique to each chemical (Imanishi and Harada, 2004) 2D-MS Microcystin Mouse liver PP1-NIPP1 complex ID as microcystin binding target (Izzotti et al., 2004) Ab array Chromium VI Rat lung DNA Array/proteomic; mRNA, protein expression coupled only during DNA damage and not homeostasis. (Jin et al., 2004) 2D-MS MNNG Human amnion FL cells 18 proteins IDed; Zn-finger family proteins altered. "
[Show abstract][Hide abstract] ABSTRACT: Global measurement of proteins and their many attributes in tissues and biofluids defines the field of proteomics. Toxicoproteomics, as part of the larger field of toxicogenomics. seeks to identify critical proteins and pathways in biological systems that are affected by and respond to adverse chemical and environmental exposures using global protein expression technologies. Toxicoproteomics integrates 3 disciplinary areas: traditional toxicology and pathology, differential protein and gene expression analysis, and systems biology. Key topics to be reviewed are the evolution of proteomics, proteomic technology platforms and their capabilities with exemplary studies from biology and medicine, a review of over 50 recent studies applying proteomic analysis to toxicological research, and the recent development of databases designed to integrate -Omics technologies with toxicology and pathology. Proteomics is examined for its potential in discovery of new biomarkers and toxicity signatures, in mapping serum,plasma. and other biofluid proteomes, and in parallel proteomic and transcriptomic studies. The new field of toxicoproteomics is uniquely positioned toward an expanded understanding of protein expression during toxicity and environmental disease for the advancement of public health.
"Ganoderma lucidum extracts in mouse spleen cells [Wang et al., 2002]. Animal models have also been directly utilized to study the drug mechanisms such as the inflammatory effects stimulated by interleukin and interferon in rat trigeminal ganglia [Friso et al., 2001], insulin processing mediated by insulin sensitizer drug rosiglitazone in pancreatic islets of obese mice [Sanchez et al., 2002] and the actions of 2,3,7,8-tetrachlorodibenzo-p-dioxin in rats [Bruno et al., 2002]. "
[Show abstract][Hide abstract] ABSTRACT: Proteomics is a research field aiming to characterize molecular and cellular dynamics in protein expression and function on a global level. The introduction of proteomics has been greatly broadening our view and accelerating our path in various medical researches. The most significant advantage of proteomics is its ability to examine a whole proteome or sub-proteome in a single experiment so that the protein alterations corresponding to a pathological or biochemical condition at a given time can be considered in an integrated way. Proteomic technology has been extensively used to tackle a wide variety of medical subjects including biomarker discovery and drug development. By complement with other new technique advances in genomics and bioinformatics, proteomics has a great potential to make considerable contribution to biomarker identification and to revolutionize drug development process. This article provides a brief overview of the proteomic technologies and their application in biomarker discovery and drug development.
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