Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Complementary proteomics and transcriptomics sheds light on drug detoxifying proteins of the endoplasmic reticulum

ABSTRACT: The hepatic endoplasmic reticulum contains a series of enzymes that oxidize and conjugate lipid and steroids. Together, these enzymes form a molecular machine that plays key roles in the metabolism of both endogenous and xenobiotic compounds. To characterize this molecular machine, we used quantitative proteomics to assess the frequency of occurrence of detected peptides within each primary sequence, leading to the assessment of the relative abundances of 137 of these proteins. These 137 proteins include over 38 different cytochrome P450s, 11 glucuronosyltransferases, and 9 carboxylesterases. Our sensitive approach was able to detect P450 allelic isoforms which differ by only a single amino acid and clearly resolved 4 splice variants of the glucuronosyltransferases. We identified a cytosolically-exposed DID motif for 3 cytochrome P450s that were located with high abundance in the Golgi apparatus as well the lack of a C-terminal HXEL motif for the sole carboxylesterase highly abundant in the Golgi. Using gene expression microarrays, we also characterized the hepatic transcriptome, and a comparison of proteomics and transcriptomics indicated a requirement for both technologies in order to provide insight into protein families of drug detoxification. In this way, a major hurdle of hepatotoxicity related to drug development may be overcome. Keywords: steady-state mRNA levels

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE9226 | GEO | 2009/11/24

SECONDARY ACCESSION(S): PRJNA102823

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Similar Datasets

Transcription profiling of rat live from Sprague Dawley animals - a complementary proteomics and transcriptomics approach sheds light on drug detoxifying proteins of the endoplasmic reticulum

Project description:The hepatic endoplasmic reticulum contains a series of enzymes that oxidize and conjugate lipid and steroids. Together, these enzymes form a molecular machine that plays key roles in the metabolism of both endogenous and xenobiotic compounds. To characterize this molecular machine, we used quantitative proteomics to assess the frequency of occurrence of detected peptides within each primary sequence, leading to the assessment of the relative abundances of 137 of these proteins. These 137 proteins include over 38 different cytochrome P450s, 11 glucuronosyltransferases, and 9 carboxylesterases. Our sensitive approach was able to detect P450 allelic isoforms which differ by only a single amino acid and clearly resolved 4 splice variants of the glucuronosyltransferases. We identified a cytosolically-exposed DID motif for 3 cytochrome P450s that were located with high abundance in the Golgi apparatus as well the lack of a C-terminal HXEL motif for the sole carboxylesterase highly abundant in the Golgi. Using gene expression microarrays, we also characterized the hepatic transcriptome, and a comparison of proteomics and transcriptomics indicated a requirement for both technologies in order to provide insight into protein families of drug detoxification. In this way, a major hurdle of hepatotoxicity related to drug development may be overcome. Experiment Overall Design: Four replicate Sprague-Dawley rats were assessed, each on an individual microarray.

2009-11-28 | E-GEOD-9226 | biostudies-arrayexpress

Effect of Traumatic Brain Injury, Erythropoietin and Anakinra on Hepatic Metabolizing Enzymes and Transporters in an Experimental Rodent Model

Project description:In contrast to the considerable in vitro and in vivo data demonstrating a decrease in cytochrome P450 (CYP) activity in inflammation and infection, clinically, traumatic brain injury (TBI) results in an increase in CYP and UDP glucuronosyltransferases (UGT) activity. The objective of this study was to determine the effects of TBI alone and along with treatment with either erythropoietin (EPO) or anakinra on gene expression of hepatic inflammatory proteins and drug metabolizing enzymes and transporters in a cortical contusion impact (CCI) injury animal model. Microarray-based transcriptional profiling was used to determine the effect on gene expression at 24 h, 72 h and 7 days post-CCI.

2015-06-01 | GSE64886 | GEO

Transcription profiling of limb buds from E12.5 wild type and POR conditional knock out mice

Project description:POR is the obligate electron donor for all microsomal P450 enzymes. POR was conditionally knocked out during limb development at E9.5. The experiment was set up to answer two questions. First, to investigate which genes are differentially expressed in POR deficient limb buds and are influenced by a metabolically inactive cytochrome P450 system. Second, to assess which P450 enzymes are expressed at all at E12.5 during limb development and therefore which metabolic pathways involving this group of enzymes are present at this stage. Differentially expressed genes were analysed by comparing 5 wild type control samples with 5 conditional knock out samples (whole forelimb buds each). Present calls of control samples were screened for annotated P450s to establish a list of expressed P450 enzymes at E12.5.

2008-10-31 | E-TABM-367 | biostudies-arrayexpress

Transcription profiling of mouse models with liver-specific deletion or global hypomorphic expression of the NADPH-cytochrome P450 reductase gene to study molecular mechanisms for the regulation of microsomal cytochrome P450 and the fatty liver phenotype.

Project description:NADPH-cytochrome P450 reductase (CPR) is important for the functions of many enzymes, such as microsomal cytochrome P450 (P450) monooxygenases and heme oxygenases. Two mouse models with deficient CPR expression in adults were recently generated in this laboratory: liver-Cpr-null (with liver-specific Cpr deletion) (Gu et al., J. Biol. Chem., 278, 25895-25901, 2003) and Cpr-low (with reduced CPR expression in all organs examined) (Wu et al. J. Pharmacol. Expt. Ther. 312, 35-43, 2005). The phenotypes included a reduced serum cholesterol level and an induction of hepatic P450 in both models, and hepatomegaly and fatty liver in the liver-Cpr-null mouse alone. Our aim was to identify hepatic gene-expression changes related to these phenotypes. Cpr-lox mice, which have normal CPR expression (Wu et al., Genesis, 36, 177-181, 2003.), were used as the control in microarray analysis. A detailed analysis of the gene-expression changes in lipid metabolism and transport pathways revealed potential mechanisms, such as an increased activation of constitutive androstane receptor (CAR) and a decreased activation of peroxisomal proliferators activated receptor alpha (PPARï?¡) by precursors of cholesterol biosynthesis, that underlie common changes (e.g., induction of multiple P450s and inhibition of genes for fatty acids metabolism) in response to CPR-loss in the two mouse models. Moreover, we also uncovered model-specific gene-expression changes, such as the induction of a lipid translocase (CD36 antigen) and the suppression of carnitine O-palmitoyltransferase 1 (CPT1a) and acyl-CoA synthetase long-chain family member 1 (Acsl1), that are potentially responsible for the severe hepatic lipidosis observed in liver-Cpr-null, but not Cpr-low mice.

2007-08-31 | E-GEOD-2362 | biostudies-arrayexpress

Transcriptomic analyses of different Tetranychus urticae strains that differ in their resistance to the acaricide pyflubumide

Project description:No major transcriptomic differences were observed between JPR-R1/2 and JPR when strains were not exposed to pyflubumide. Only three genes (tetur13g01730, tetur14g03160, and tetur14g01700) were consistently up-regulated in JPR-R1/2, but none coded for enzymes that are known to be associated with xenobiotic metabolism. As a next step, we used the original and susceptible JPO strain, the parent of JPR, as a reference to investigate transcriptomic changes. Compared to JPO, 16 cytochrome P450s were up-regulated in JPR, whereas two CCE genes were down-regulated. In both JPR-R1/2, 13 cytochrome P450s were up-regulated compared to JPO, of which 11 genes already showed significant up-regulation in JPR (CYP392D2, CYP392D6, and CYP392E8 had the highest average transcriptional increase). As only limited constitutive gene expression differences were observed between JPR and the JPR-R1/2 strains, the plastic, environmentally dependent, transcriptomic changes were characterized to shed light on the molecular basis of pyflubumide resistance. After nine hours of exposure to a non-lethal dose of pyflubumide (100 mg/L), no DEGs were detected in JPR-R1 upon pyflubumide exposure, whereas only a single DEG (tetur04g04350, log2FC of 1.17) was observed in JPR. After 24 hours of exposure to pyflubumide (100mg/L), large transcriptomic responses were observed in JPR-R1/2. Eight cytochrome P450s were part of the positively correlated response, of which five (CYP385C4, CYP385C4v2, CYP392D2, CYP392E8, and CYP392A4) showed up-regulation after exposure to pyflubumide.

2020-10-10 | GSE138192 | GEO

High-fat diet induces the modulation of hepatic nuclear receptors and their target genes in C57BL/6 mice

Project description:We reported the hepatic gene expression profiling in mice fed with a high fat diet compared to the controls. We found that the modulation of pathways related to peroxisome proliferator-activated receptors, cytochrome P450s, and ATP-binding cassette transporters in high fat diet mice. Particularly, constitutive androstane receptor and pregnane X receptor signature genes Cyp2b10 and Cyp3a11 were significantly increased in high fat diet mice compared to controls. Liver X receptor mediated Abcg5 and Abcg8 were also found increased by HFD treatment.

2018-08-28 | GSE118070 | GEO

Altered hepatic gene expression in liver-Cpr-null and Cpr-low mice

Project description:NADPH-cytochrome P450 reductase (CPR) is important for the functions of many enzymes, such as microsomal cytochrome P450 (P450) monooxygenases and heme oxygenases. Two mouse models with deficient CPR expression in adults were recently generated in this laboratory: liver-Cpr-null (with liver-specific Cpr deletion) (Gu et al., J. Biol. Chem., 278, 25895–25901, 2003) and Cpr-low (with reduced CPR expression in all organs examined) (Wu et al. J. Pharmacol. Expt. Ther. 312, 35-43, 2005). The phenotypes included a reduced serum cholesterol level and an induction of hepatic P450 in both models, and hepatomegaly and fatty liver in the liver-Cpr-null mouse alone. Our aim was to identify hepatic gene-expression changes related to these phenotypes. Cpr-lox mice, which have normal CPR expression (Wu et al., Genesis, 36, 177-181, 2003.), were used as the control in microarray analysis. A detailed analysis of the gene-expression changes in lipid metabolism and transport pathways revealed potential mechanisms, such as an increased activation of constitutive androstane receptor (CAR) and a decreased activation of peroxisomal proliferators activated receptor alpha (PPAR-gamma) by precursors of cholesterol biosynthesis, that underlie common changes (e.g., induction of multiple P450s and inhibition of genes for fatty acids metabolism) in response to CPR-loss in the two mouse models. Moreover, we also uncovered model-specific gene-expression changes, such as the induction of a lipid translocase (CD36 antigen) and the suppression of carnitine O-palmitoyltransferase 1 (CPT1a) and acyl-CoA synthetase long-chain family member 1 (Acsl1), that are potentially responsible for the severe hepatic lipidosis observed in liver-Cpr-null, but not Cpr-low mice. Keywords = Cytochrome P450 Keywords = NADPH-cytochrome P450 reductase Keywords = transgenic mice Keywords = liver Keywords = nuclear receptor Keywords: other

2005-07-16 | GSE2362 | GEO

Cytochrome P450s protect against oxidative damage via a preferred orientation of the heme cofactor

Project description:Cytochrome P450 enzymes (P450s) are a superfamily of monooxygenases that utilise a cysteine thiolate ligated heme moiety to perform a wide range of demanding oxidative transformations. Given the oxidative power of active intermediate formed within P450s during their active cycle, it is remarkable that these enzymes can avoid self-oxidation as well as retaining the axial cysteine ligand in the deprotonated – and thus highly acidic – thiolate form. Whilst little is known about the process of heme incorporation during P450 folding, there is an overwhelming preference for one heme orientation within the P450 active site. Indeed, very few structures to date contain an alternate heme orientation, of which two are OxyA homologues from glycopeptide antibiotic (GPA) biosynthesis. Given the apparent preference for the unusual heme orientation shown by OxyA enzymes, we investigated the OxyA homologue from kistamicin biosynthesis, which is an atypical GPA. We determined that OxyAkis is highly sensitive to oxidative damage by peroxide, with both UV an EPR measurements shows a rapid bleaching of the heme signal. We determined the structure of OxyAkis and found a mixed population of heme orientations present in this enzyme. Analysis further revealed that an unprecedented oxidative modification of the heme was detected, which that correlated with the presence of the alternate heme orientation in the protein. These results provide evidence that the typical heme orientation in Cytochrome P450s can help to prevent potential autocatalytic modification of the heme – and hence deactivation of the enzyme – during P450 catalysis. It also suggests that some P450 enzymes involved in GPA biosynthesis may be especially prone to oxidative damage due to the heme orientation found in their active sites.

2022-05-20 | PXD030867 | Pride

Identification of human genes of which expression are induced by 4MU-xyloside treatment

Project description:The Golgi stress response is a homeostatic mechanism that augments the functional capacity of the Golgi apparatus when Golgi function becomes insufficient (Golgi stress). Three response pathways of the Golgi stress response have been identified in mammalian cells, the TFE3, HSP47 and CREB3 pathways, which augment the capacity of specific Golgi functions such as N-glycosylation, anti-apoptotic activity and pro-apoptotic activity, respectively. On the contrary, glycosylation of proteoglycans (PGs) is another important function of the Golgi, although the response pathway upregulating expression of glycosylation enzymes for PGs in response to Golgi stress remains unknown. Here, we found that expression of glycosylation enzymes for PGs was induced upon insufficiency of PG glycosylation capacity in the Golgi (PG-Golgi stress), and that transcriptional induction of genes encoding glycosylation enzymes for PGs was independent of the known Golgi stress response pathways and ER stress response. Promoter analyses of genes encoding these glycosylation enzymes revealed the novel enhancer element PGSE, which regulates their transcriptional induction upon PG-Golgi stress. From these observations, the response pathway we discovered is a novel Golgi stress response pathway, which we have named the PG pathway.

2018-08-01 | GSE117938 | GEO

Identification of human gene expression induced by 4MU-xyloside treatment

2018-09-14 | GSE119909 | GEO

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data