Proteomics

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Acute to prolonged lif cycle proteomic profiling of heat shock SGs


ABSTRACT: Time-resolved proteomic profiling of SGs from cultured human U2OS cells over a prolonged heat-shock (HS) and a successive recovery period, sampling at the 6 time points of HS exposure (10, 20, 30, 60, 120, 180 min), as well as 2 time points of recovery (5, 10 min). From the sampled cells, we prepared SG samples using a differential centrifugation plus endogenous G3BP1 immunoprecipitation protocol. Peptides were subsequently TMT-labeled and analyzed by quantitative LC-MS/MS. In G3BP1 dataset, rep1-rep3, HS-10, 20, 30, 60, 120, 180, re5, re10, SG-ls, G3BP1-IP samples were labeled with TMT-126, 127N, 127C, 128N, 128C,129N, 129C, 130N, 130C, 131 label Reagents, respectively; G3BP1_HS0 was a dataset created to integrate a non-heat shock control (HS-0) into the G3BP1 dataset, contains HS0, HS10, and HS60, and the results was integrated into the G3BP1 dataset using the ComBat processing, HS-0_rep1, HS-0_rep2, HS-0_rep3, HS-10_rep1, HS-10_rep2, HS-10_rep3, HS-60_rep1, HS-60_rep2, HS-60_rep3 samples were labeled with TMT-126, 127N, 127C, 128N, 128C,129N, 129C, 130N, 130C label Reagents, respectively. In CAPRIN1 dataset, rep1-rep3, HS-0, 10, 20, 30, 60, 120, 180, re5, re10 samples were labeled with TMT-126, 127N, 127C, 128N, 128C,129N, 129C, 130N, 130C label Reagents, respectively.

INSTRUMENT(S): Q Exactive HF

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

SUBMITTER: Shuyao Hu  

PROVIDER: MSV000092790 | MassIVE | Wed Aug 30 07:30:00 BST 2023

SECONDARY ACCESSION(S): PXD044967

REPOSITORIES: MassIVE

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Publications

Time-resolved proteomic profiling reveals compositional and functional transitions across the stress granule life cycle.

Hu Shuyao S   Zhang Yufeng Y   Yi Qianqian Q   Yang Cuiwei C   Liu Yanfen Y   Bai Yun Y  

Nature communications 20231127 1


Stress granules (SGs) are dynamic, membrane-less organelles. With their formation and disassembly processes characterized, it remains elusive how compositional transitions are coordinated during prolonged stress to meet changing functional needs. Here, using time-resolved proteomic profiling of the acute to prolonged heat-shock SG life cycle, we identify dynamic SG proteins, further segregated into early and late proteins. Comparison of different groups of SG proteins suggests that their biochem  ...[more]

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