Dataset Information

A Tandem Mass Tag (TMT) proteomic analysis during the early phase of experimental pancreatitis

ABSTRACT: We performed in a rat pancreatitic model a quantitative shotgun proteomic profiling of the soluble and whole membrane fractions from the pancreas during the early phase of cerulein (Cer)-induced AP compared to controls. Three control and three pancreatitic samples from both the soluble and the whole membrane fractions were analysed in two independent TMT6plex experiments. After trypsin digestion and TMT labelling, the peptide mixtures were fractionated into 12 fractions by off-gel electrophoresis. ESI LTQ-OT MS was performed on a LTQ Orbitrap velos equipped with a NanoAcquity system from Waters. MS data were analysed for protein identifications using EasyProt (v2.3) and peak lists were generated into .mgf format with EasyProtConv (CID/HCD merging was used to improve peptide identification and quantification). Searches were conducted against UniProt Swiss-Prot database (UniProtKB release 2014_10 of Oct, 29, 2014), specifying Rattus norvegicus taxonomy. Only proteins with at least two unique peptide sequences and a false discovery rate (FDR) ≤ 1 % were selected for further quantification. Proteins were clustered based on shared peptides indistinguishable by MS. Quantification was conducted using Isobar R package (v.1.9.3.2). We identified 997 proteins, of which 353 were significantly differentially expressed (22, 256 or 55 in both, the soluble or the membrane fractions, respectively).

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Pancreas

DISEASE(S): Acute Pancreatitis

SUBMITTER: Violeta García-Hernández

LAB HEAD: Jesús Sánchez-Yagüe

PROVIDER: PXD007096 | Pride | 2018-05-22

REPOSITORIES: Pride

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Publications

A tandem mass tag (TMT) proteomic analysis during the early phase of experimental pancreatitis reveals new insights in the disease pathogenesis.

García-Hernández Violeta V Sánchez-Bernal Carmen C Schvartz Domitille D Calvo José J JJ Sanchez Jean-Charles JC Sánchez-Yagüe Jesús J

Journal of proteomics 20180417

Changes in the protein expression occurring within the initiation phase of acute pancreatitis (AP) might be vital in the development of this complex disease. However, the exact mechanisms involved in the onset of AP remains elusive and most of our knowledge about the pathobiology of AP comes from animal models. We performed in a rat pancreatitic model a high-throughput shotgun proteomic profiling of the soluble and whole membrane fractions from the pancreas during the early phase of cerulein (Ce ...[more]

PMID: 29678717

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Project description:modENCODE_submission_2759 This submission comes from a modENCODE project of Steven Henikoff. For full list of modENCODE projects, see http://www.genome.gov/26524648 Project Goal: We applied genome-wide profiling to successive salt-extracted fractions of micrococcal nuclease-treated Drosophila chromatin. Chromatin fractions extracted with 80 mM or 150 mM NaCl after digestion contain predominantly mononucleosomes and represent classical "active" chromatin. Profiles of these low-salt soluble fractions display phased nucleosomes over transcriptionally active genes that are locally depleted of histone H3.3 and correspond closely to profiles of histone H2Av (H2A.Z) and RNA polymerase II. This correspondence suggests that transcription can result in loss of H3.3+H2Av nucleosomes and generate low-salt soluble nucleosomes. Nearly quantitative recovery of chromatin is obtained with 600 mM NaCl; however, the remaining insoluble chromatin is enriched in actively transcribed regions. Salt-insoluble chromatin likely represents oligonucleosomes that are attached to large protein complexes. Both low-salt extracted and insoluble chromatin are rich in sequences that correspond to epigenetic regulatory elements genome-wide. The presence of active chromatin at both extremes of salt solubility suggests that these salt fractions capture bound and unbound intermediates in active processes, thus providing a simple, powerful strategy for mapping epigenome dynamics. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf EXPERIMENT TYPE: tiling array analysis. BIOLOGICAL SOURCE: Cell Line: Kc167; Tissue: embryo-derived cell-line; Developmental Stage: late embryonic stage; Genotype: se/e; Sex: Female; NUMBER OF REPLICATES: 2; EXPERIMENTAL FACTORS: Cell Line Kc167; biochemical fraction (extract) soluble fraction; sodium chloride concentration (Compound) 600 mM; extraction time (sampling_time_point) 1 to 2 hours

Dataset Information

A Tandem Mass Tag (TMT) proteomic analysis during the early phase of experimental pancreatitis

Publications

A tandem mass tag (TMT) proteomic analysis during the early phase of experimental pancreatitis reveals new insights in the disease pathogenesis.

Similar Datasets

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