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ATR phosphoproteomics screening using berzosertib and gartisertib

ABSTRACT: ATR phosphoproteomics screening using berzosertib and gartisertib Quantitiative phosphoproteomics using TMT10plex labelling and basic reverse phase HPLC prefractionation. Experimental and data analysis procedures detailed in the accompanying manuscript. Data analysis scripts included in this submission.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: John Rouse

PROVIDER: PXD040469 | JPOST Repository | Mon Jan 15 00:00:00 GMT 2024

REPOSITORIES: jPOST

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			Action	DRS
	20180503_FW011_TMT-FRACTION_01-1_FW_180504003709.raw	Raw
	20180503_FW011_TMT-FRACTION_01-2_FW.raw	Raw
	20180503_FW011_TMT-FRACTION_01-3_FW.raw	Raw
	20180503_FW011_TMT-FRACTION_02-1_FW.raw	Raw
	20180503_FW011_TMT-FRACTION_02-2_FW.raw	Raw

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Project description:Pilotexp_IMAC_TiO2_eluate_ATM_ATR_dep_phos_15min_postIR: Proteins were isolated from 4 different conditions of Arabidopsis thaliana plants: wild type untreated, wild type irradiated, atm/atr mutant untreated, atm/atr mutant irradiated The proteins were tryptically digested and labelled each with one channel ofthe iTRAQ4plex reagent. The samples were mixed and subjected to phosphopeptide enrichment by IMAC followed by TiO2. The enriched phosphopeptides were separated by SCX and each fraction was analysed by nano-reversed phase HPLC coupled online to a LTQ Orbitrap Velos. Each peptide was fragmented both by CID (MSA) and HCD, using the iontrap-MS analyser for CID and the Orbitrap detector for HCD scans. IMAC_TiO2_eluate_ATM_ATR_dep_phos_15min_postIR: Proteins were isolated from 4 different conditions of Arabidopsis thaliana plants: wild type untreated, wild type irradiated, atm/atr mutant untreated, atm/atr mutant irradiated The proteins were tryptically digested and labelled each with one channel ofthe iTRAQ4plex reagent. The samples were mixed and subjected to phosphopeptide enrichment by IMAC followed by TiO2. The enriched phosphopeptides were separated by SCX and each fraction was analysed by nano-reversed phase HPLC coupled online to a LTQ Orbitrap Velos. Each peptide was fragmented both by CID (MSA) and HCD, using the iontrap-MS analyser for CID and the Orbitrap detector for HCD scans. IMAC_TiO2_FT_ATM_ATR_dep_phos_15min_postIR: Proteins were isolated from 4 different conditions of Arabidopsis thaliana plants: wild type untreated, wild type irradiated, atm/atr mutant untreated, atm/atr mutant irradiated The proteins were tryptically digested and labelled each with one channel ofthe iTRAQ4plex reagent. The samples were mixed and subjected to phosphopeptide enrichment by IMAC followed by TiO2. The unphosphorylated peptides found in the flowthrough of the TiO2 column were separated by SCX and each fraction was analysed by nano-reversed phase HPLC coupled online to a LTQ Orbitrap Velos. Each peptide was fragmented both by CID and HCD, using the iontrap-MS analyser for CID and the Orbitrap detector for HCD scans. The iTRAQ ratio obtained for the unmodified peptides from this flowthrough sample was used to calculate protein ratios for the 4 different conditions.

Dataset Information

ATR phosphoproteomics screening using berzosertib and gartisertib

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