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Human “free-N-terminome” analysis

ABSTRACT: The high throughput characterization of protein N-termini is becoming an emerging challenge in the proteomics and proteogenomics fields. The present study describes the N-terminal oriented analysis of human mitochondria enriched samples by the trimethoxyphenyl phosphonium (TMPP) labeling approach (N-TOP and dN-TOP approaches (Gallien et al, 2009, Bertaccini et al. 2013)). The dN-TOP strategy based on a double light/heavy TMPP labeling has been optimized in order to improve and automate the workflow for efficient, fast and reliable high throughput N-terminome analysis.

INSTRUMENT(S): maXis 4G, amaZon ETD

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Dr Alain Van Dorsselaer

PROVIDER: MSV000080204 | MassIVE |

SECONDARY ACCESSION(S): PXD001521

REPOSITORIES: MassIVE

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N-terminome analysis of the human mitochondrial proteome.

Vaca Jacome Alvaro Sebastian AS Rabilloud Thierry T Schaeffer-Reiss Christine C Rompais Magali M Ayoub Daniel D Lane Lydie L Bairoch Amos A Van Dorsselaer Alain A Carapito Christine C

Proteomics 20150608 14

The high throughput characterization of protein N-termini is becoming an emerging challenge in the proteomics and proteogenomics fields. The present study describes the free N-terminome analysis of human mitochondria-enriched samples using trimethoxyphenyl phosphonium (TMPP) labelling approaches. Owing to the extent of protein import and cleavage for mitochondrial proteins, determining the new N-termini generated after translocation/processing events for mitochondrial proteins is crucial to unde ...[more]

PMID: 25944712

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Human “free-N-terminome” analysis

Project description:The high throughput characterization of protein N-termini is becoming an emerging challenge in the proteomics and proteogenomics fields. The present study describes the N-terminal oriented analysis of human mitochondria enriched samples by the trimethoxyphenyl phosphonium (TMPP) labeling approach (N-TOP and dN-TOP approaches (Gallien et al, 2009, Bertaccini et al. 2013)). The dN-TOP strategy based on a double light/heavy TMPP labeling has been optimized in order to improve and automate the workflow for efficient, fast and reliable high throughput N-terminome analysis.

2015-06-02 | PXD001521 | Pride

Human “free-N-terminome” analysis

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Comparison of Labeling Methods for gene expression analysis on the NimbleGen platform

Project description:Comparison of ds-cDNA, Indirect and Direct Random Labeling Methods for gene expression analysis on the NimbleGen platform. Expression profiles from artemisinic acid-producing S. cerevisiae strain EPY330 and non-producing strain EPY338 are compared for each labeling method tested. The labeling methods and their comparison are described in detail in Ouellet et al., BMC Biotech 2009. Strains were described in detail previously in Ro et al. BMC Biotechnol 2008, 8(1):83 [PMID: 18983675]

2009-11-25 | GSE15816 | GEO

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Project description:adt12-01_uprt - plant rabt n°1 - Impact of RNA labeling by Plant RABT on plant transcriptome - Determine the incidence of RNA marking by the RABT method Plantation on the plants transcriptome. In several studies Clearly et al. have described a method referred as "4TU tagging" which can be use to study mRNA synthesis and decay either in a mixed population of cells or in a specific cell type (Cleary, Meiering et al. 2005, Zeiner, Cleary et al. 2008, Miller, Robinson et al. 2009, Rabani et al., 2011). Through the specific cell type expression in drosophila and mammalian cells of theToxoplasma gondii uracil PhosphoribosylTransferase activity and the metabolization of the uracil analog 4-thiouracil (4 TU), mRNA were selectively tagged, purified and used in microarray based analysis.

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Streptococcus agalactiae temperature changes

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Argonaute HITS-CLIP in wild-type and APOBEC1-deficient BMDMs

Project description:To investigate the possible consequences of APOBEC1 editing for miRNA targeting, high-throughput sequencing of RNA isolated by cross-linking immunoprecipitation (HITS-CLIP) of the Argonaute (Ago) proteins (Chi et al., 2009) was performed in BMDMs derived from wild-type and Apobec1-/- littermates.

2017-11-21 | GSE58798 | GEO

Dynamic transcriptome analysis of osmotic stress response

Project description:Dynamic transcriptome analysis of osmotic stress response. Unperturbed RNA synthesis and decay rates can be obtained via metabolic RNA labeling and kinetic modeling (Cleary et al, 2005; Kenzelmann et al, 2007; Dolken et al, 2008; Friedel and Dolken, 2009; Miller et al, 2009). The nucleoside analog 4-thiouridine (4sU) is taken up by eukaryotic cells and incorporated into mRNA during Pol II transcription (Melvin et al, 1978). The thiol-labeled newly transcribed RNA can then be isolated by affinity chromatography (Kenzelmann et al, 2007) or by biotinylation and purification with streptavidin-coated magnetic beads (Cleary et al, 2005; Dolken et al, 2008).

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2019-10-01 | GSE88839 | GEO

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