Dataset Information

K562 Elutriation cell cycle phase fractions 2D-MS-CETSA

ABSTRACT: K562 cells were synchronized by cell size difference based Elutriation scheme into G1, S and G2 fractions. Three biological replicates were collected on different days, and assembled into a 2D-CETSA scheme with 6 different heating temperatures (37ºC, 47ºC, 50ºC, 52ºC,54ºC and 57ºC), (Figure 1A, Methods section of the associated paper), with each set of samples heated at one temperature included in the same isobaric TMT10 set and measured at the same time on Mass Spectrometer. The labeling order of samples in the TMT10 channels is as follows: 37C/47C samples: Biorep1_G1(126), Biorep1_S(127N), Biorep1_G2(127C), Biorep2_G1(128N), Biorep2_S(128C), Biorep2_G2(129N), Biorep3_S(129C), Biorep3_G1(130N), Biorep3_G2(130C), mixed_control(131); 50C/52C/54C/57C samples: Biorep1_G1(126), Biorep1_S(127N), Biorep1_G2(127C), Biorep2_G1(128N), Biorep2_S(128C), Biorep2_G2(129N), Biorep3_G1(129C), Biorep3_S(130N), Biorep3_G2(130C), mixed_control(131);

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Pär Nordlund

PROVIDER: PXD009171 | JPOST Repository | Tue May 08 00:00:00 BST 2018

REPOSITORIES: jPOST

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20170414_LY_TY_Elutri_54C_3bio_rep1_1.raw	Raw
	20170414_LY_TY_Elutri_54C_3bio_rep1_10.raw	Raw
	20170414_LY_TY_Elutri_54C_3bio_rep1_11.raw	Raw
	20170414_LY_TY_Elutri_54C_3bio_rep1_12.raw	Raw
	20170414_LY_TY_Elutri_54C_3bio_rep1_13.raw	Raw

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Publications

Modulation of Protein-Interaction States through the Cell Cycle.

Dai Lingyun L Zhao Tianyun T Bisteau Xavier X Sun Wendi W Prabhu Nayana N Lim Yan Ting YT Sobota Radoslaw M RM Kaldis Philipp P Nordlund Pär P

Cell 20180426 6

Global profiling of protein expression through the cell cycle has revealed subsets of periodically expressed proteins. However, expression levels alone only give a partial view of the biochemical processes determining cellular events. Using a proteome-wide implementation of the cellular thermal shift assay (CETSA) to study specific cell-cycle phases, we uncover changes of interaction states for more than 750 proteins during the cell cycle. Notably, many protein complexes are modulated in specifi ...[more]

PMID: 29706543

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Project description:The small molecule YC-1 was identified as identified as a selectively active agent against subsets of the two major live cancer subtypes, intrahepatic cholangiocarcinoma and hepatocellular carcinoma. Response to YC-1 is dependent on the expression of the sulfotransferase SULT1A1, which activates YC-1 through sulfonation. The contributions of mass spectrometry to the study were: (i) identifying SULT1A1 as the key player in YC-1 response through observing the protein in a cell line with acquired resistance to the small molecule treatment, (ii) corroborating the role of SULT1A1 in the YC-1 activity by correlating protein expression profiles and drug responses across 37 biliary cancer cell lines, (iii) showing that SULT1A1 expression was associated with IDH mutations, FGFR2 fusion, and BAP1 loss in vivo studying patient-derived xenograft (PDX) models, and (iv) identifying protein targets in a protein enrichment experiment with an immobilized drug. (i) and (ii): Cell line samples from datasets (i) and (ii) were analyzed together in a set of 12 TMT sets (ShiBardeesy_CellLines_TMT1 to TMT12) using TMT11-barcoding. The samples were analyzed on an Orbitrap Lumos mass spectrometer using the SPS-MS3 method. Prior to analysis, samples were fractionated offline using high pH reversed chromatography (12 fractions per TMT set). Across all TMT sets, channel 131c was used as the bridge channel (pooled digests of all samples). Samples were labeled as follows (rep = replicate: (i) Cell lines samples with acquired resistance to YC-1 treatment: ShiBardeesy_CellLines_TMT1: RBE_rep_1 (126), RBE YC1 Resistant 6_rep_1 (129c). ShiBardeesy_CellLines_TMT2: RBE YC1 Resistant 3_rep_1 (129n), RBE YC1 Resistant 1_rep_1 (130n). ShiBardeesy_CellLines_TMT3: RBE YC1 Resistant 2_rep_1 (126), RBE YC1 Resistant 5_rep_1 (130c). ShiBardeesy_CellLines_TMT4: RBE YC1 Resistant 4_rep_1 (126). ShiBardeesy_CellLines_TMT5: RBE YC1 Resistant 1_rep_2 (127n), RBE YC1 Resistant 4_rep_2 (128c), RBE YC1 Resistant 3_rep_2 (130c). ShiBardeesy_CellLines_TMT6: RBE YC1 Resistant 2_rep_2 (126), RBE YC1 Resistant 6_rep_2 (127c). ShiBardeesy_CellLines_TMT7: RBE_rep_2 (128n), RBE YC1 Resistant 5_rep_2 (130n). (ii) Proteomes of untreated biliary cell lines (baseline): ShiBardeesy_CellLines_TMT1: RBE_rep_1 (126), GB2_rep_1 (127n), HuCCT1_rep_1 (127c), ECC3_rep_1 (128n), ICC10_rep_1 (128c), KMCH-1_rep_1 (129n), SNU-1196_rep_1 (130n). ShiBardeesy_CellLines_TMT2: ICC13-7_rep_1 (126), ICC8_rep_1 (127n), ICC9_rep_1 (127c), SNU-478_rep_1 (128n), CC-SW-1_rep_1 (128c), CC-LP-1_rep_1 (129c), SSP-25_rep_1 (130c), ICC10-8_rep_1 (131n) ShiBardeesy_CellLines_TMT3: SNU-1079_rep_1 (127n), ICC5_rep_1 (127c), ICC7_rep_1 (128c), COR-L105_rep_1 (129n), ICC17_rep_1 (129c), GBC1_rep_1 (131n). ShiBardeesy_CellLines_TMT4: ICC3_rep_1 (127n), ICC2_rep_1 (127c), ICC106_rep_1 (128n), ICC11_rep_1 (128c), HKGZ-CC_rep_1 (129n), SNU-869_rep_1 (129c), HuH28_rep_1 (130n), ICC12_rep_1 (130c), SNU-308_rep_1 (131n). ShiBardeesy_CellLines_TMT5: ICC2_rep_2 (126), ECC3_rep_2 (127c), GB2_rep_2 (128n), ICC11_rep_2 (129n), ICC7_rep_2 (129c), SNU-869_rep_2 (130n), SSP-25_rep_2 (131n). ShiBardeesy_CellLines_TMT6: CC-LP-1_rep_2 (127n), COR-L105_rep_2 (128n), HuCCT1_rep_2 (129n), ICC12_rep_2 (129c), ICC3_rep_2 (131n), ICC5_rep_2 (130n), SNU-478_rep_2 (130c). ShiBardeesy_CellLines_TMT7: ICC10_rep_2 (126), ICC10-6_rep_2 (127n), ICC10-8_rep_2 (127c), RBE_rep_2 (128n), ICC9_rep_2 (128c), ICC17_rep_2 (129n), ICC8_rep_2 (129c), CC-SW-1_rep_2 (130c), GBC1_rep_2 (131n). ShiBardeesy_CellLines_TMT8: ICC13-7_rep_2 (127c), HuH28_rep_2 (128n), HKGZ-CC_rep_2 (128c), SNU-1079_rep_2 (129n), KMCH-1_rep_2 (129c), SNU-1196_rep_2 (130n), SNU-308_rep_2 (131n). ShiBardeesy_CellLines_TMT9: SG231_rep_1 (126), KKU-100_rep_1 (127n), YSCCC_rep_1 (127c), ICC4_rep_1 (128n), TKKK_rep_1 (128c), ECC2_rep_1 (129n), KKU-M213/M214 (129c). ShiBardeesy_CellLines_TMT10: YSCCC_rep_2 (126), KKU-M213/M214_rep_2 (127n), SG231_rep_2 (127c), TKKK_rep_2 (129n), ECC2_rep_2 (129c), ICC4_rep_2 (130n), KKU-100_rep_2 (131n). ShiBardeesy_CellLines_TMT11: ICC13-7_rep_1 (129n). ShiBardeesy_CellLines_TMT12: ICC13-7_rep2 (131n)