Proteomics

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IMPRINTS-CETSA, 5-FU in parental and 5-FU-resistant HCT15 cells


ABSTRACT: Parental and 5-fluorouracil (5-FU)-resistant HCT15 cells were treated with 5-FU or vehicle for 12h and assembled into an IMPRINTS-CETSA scheme with 6 different heating temperatures (37ºC, 47ºC, 50ºC, 52ºC, 54ºC, and 57ºC) with each set of samples heated at one temperature included in the same isobaric TMT10 set and measured at the same time on the mass spectrometer. This entry contains two datasets: 1. 5-FU in parental vs. resistant HCT15 (file names “12h 5FU_par vs resHCT15”) for data referred to as HCT15_Par_5FU and HCT15_Res_5FU in the associated publication. The labelling order of samples in the TMT10 channels for this dataset was as follows: 37C/47C/50C/52C/54C/57C samples: Biorep1_Parental_Vehicle (126), Biorep1_Parental_5FU_100µM (127N), Biorep1_Resistant_5FU_16µM (127C), Biorep2_ Parental_Vehicle (128N), Biorep2_ Parental_5FU_100µM (128C), Biorep2_ Resistant_5FU_16µM (129N), Biorep3_ Parental_Vehicle (129C), Biorep3_ Parental_5FU_100µM (130N), Biorep3_ Resistant_5FU_16µM (130C), mixed_control (131). 2. 5-FU in resistant HCT15 (file names “12h 5FU_ResHCT15_diff”) for data referred to as HCT15_Res_Diff in the in the associated publication. The labelling order of samples in the TMT10 channels for this dataset was as follows: 37C/47C/50C/52C/54C/57C samples: Biorep1_ Resistant_5FU_16µM (128N), Biorep2_ Resistant_5FU_16µM (128C), Biorep3_ Resistant_5FU_16µM (129N), Biorep1_ Resistant_5FU_100µM (129C), Biorep2_ Resistant_5FU_100µM (130N), Biorep3_ Resistant_5FU_100µM (130C), mixed_control (131). For additional details see the methods section of the associated publication.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Pär Nordlund 

PROVIDER: PXD026472 | JPOST Repository | Thu Jul 15 00:00:00 BST 2021

REPOSITORIES: jPOST

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Publications

CETSA interaction proteomics define specific RNA-modification pathways as key components of fluorouracil-based cancer drug cytotoxicity.

Liang Ying Yu YY   Bacanu Smaranda S   Sreekumar Lekshmy L   Ramos Anderson Daniel AD   Dai Lingyun L   Michaelis Martin M   Cinatl Jindrich J   Seki Takahiro T   Cao Yihai Y   Coffill Cynthia R CR   Lane David P DP   Prabhu Nayana N   Nordlund Pär P  

Cell chemical biology 20210716 4


The optimal use of many cancer drugs is hampered by a lack of detailed understanding of their mechanism of action (MoA). Here, we apply a high-resolution implementation of the proteome-wide cellular thermal shift assay (CETSA) to follow protein interaction changes induced by the antimetabolite 5-fluorouracil (5-FU) and related nucleosides. We confirm anticipated effects on the known main target, thymidylate synthase (TYMS), and enzymes in pyrimidine metabolism and DNA damage pathways. However, m  ...[more]

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