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AgInsoluble

ABSTRACT: Isolation of Insoluble Aggregates from Worm Lysates

REANALYSIS of: PXD001364

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Caenorhabditis Elegans (c. Elegans)

SUBMITTER:

PROVIDER: PAe005568 | PeptideAtlas | 2015-12-31

REPOSITORIES: PeptideAtlas

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Publications

Widespread Proteome Remodeling and Aggregation in Aging C. elegans.

Walther Dirk M DM Kasturi Prasad P Zheng Min M Pinkert Stefan S Vecchi Giulia G Ciryam Prajwal P Morimoto Richard I RI Dobson Christopher M CM Vendruscolo Michele M Mann Matthias M Hartl F Ulrich FU

Cell 20150501 4

Aging has been associated with a progressive decline of proteostasis, but how this process affects proteome composition remains largely unexplored. Here, we profiled more than 5,000 proteins along the lifespan of the nematode C. elegans. We find that one-third of proteins change in abundance at least 2-fold during aging, resulting in a severe proteome imbalance. These changes are reduced in the long-lived daf-2 mutant but are enhanced in the short-lived daf-16 mutant. While ribosomal proteins de ...[more]

PMID: 25957690

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Project description:We report the isolation and sequencing of tau aggregates from [1] HEK293 cells expressing Tau-RD-P301S-CFP/YFP that have been seeded with preformed fibrils from the brain of P301S mice (B6-Tg(Thy1-MAPT*P301S)2541; referred to as Tg2541 mice). Tau aggregates were isolated by differential centrifugation followed by fluorescence automated particle sorting using a BD FACSaraia. We found that these tau aggregates were enriched for particular small non-coding RNAs, including snoRNAs and snRNAs. [2] the following mice: FvBB6F1-Tg(Camk2a-tTa),(tetO-MAPT*wt)21221 (referred to as rTg21221 or WT tau mice in the paper) and FvBB6F1-Tg(Camk2a-tTA)1Mmay, (tet)-tdTomato-Syp/EGFP)1.1Luo/J,(tetO-MAPT*P301L)4510 (referred to as rTg4510 or P301L mice in the paper). Briefly, tau aggregates were isolated by 1% sarkosyl extraction (to enrich for insoluble proteins) followed by immunoprecipitation of tau using the tau-12 antibody (see Methods section of associated paper for further details). We found that these tau aggregates were enriched for particular small non-coding RNAs, including snRNAs and some snoRNAs. [3] Sequencing of HEK293 tau biosensor cells with and without tau aggregates reveals evidence of significant splicing alterations. Specifically we observed an increase in intron retention events in cells that contain tau aggregates relative to cells without tau aggregates.

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AgTotal

Project description:Isolation of Aggregates from Worm Lysates

2015-12-31 | PAe005582 | PeptideAtlas

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AgSoluble

Project description:Isolation of soluble Aggregates from Worm Lysates

2015-12-31 | PAe005579 | PeptideAtlas