Dataset Information

Mapping physiological ADP-ribosylation using Activated Ion Electron Transfer Dissociation (AI-ETD)

ABSTRACT: ADP-ribosylation (ADPr) is a post-translational modification that plays pivotal roles in a wide range of cellular processes. Mass spectrometry (MS)-based analysis of ADPr under physiological conditions, without relying on genetic or chemical perturbation, has been hindered by technical limitations. Here, we describe the applicability of activated ion electron transfer dissociation (AI-ETD) for MS-based proteomics analysis of physiological ADPr using our unbiased Af1521 enrichment strategy. To benchmark AI-ETD, we profile 9,000 ADPr peptides mapping to >5,000 unique ADPr sites from a limited number of cells exposed to oxidative stress and identify 120% and 28% more ADPr peptides compared to contemporary strategies using ETD and electron-transfer higher-energy collisional dissociation (EThcD), respectively. Under physiological conditions, AI-ETD identifies 450 ADPr sites on low-abundant proteins, including in vivo cysteine modifications on poly(ADP-ribosyl)polymerase (PARP) 8 and tyrosine modifications on PARP14, hinting at specialist enzymatic functions for these enzymes. Collectively, our data provide insights into the physiological regulation of ADPr.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

DISEASE(S): Cervix Carcinoma

SUBMITTER: Sara Larsen

LAB HEAD: Michael Lund Nielsen

PROVIDER: PXD017417 | Pride | 2020-09-23

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20190803_LUMOS_D3K_JML-SCL-IAH_ADPr_LysC_AI-ETD15_01.raw	Raw
	20190803_LUMOS_D3K_JML-SCL-IAH_ADPr_LysC_AI-ETD15_02.raw	Raw
	20190803_LUMOS_D3K_JML-SCL-IAH_ADPr_LysC_AI-ETD15_03.raw	Raw
	20190803_LUMOS_D3K_JML-SCL-IAH_ADPr_LysC_AI-ETD15_04.raw	Raw
	20190803_LUMOS_D3K_JML-SCL-IAH_ADPr_LysC_AI-ETD20_01.raw	Raw

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Publications

Mapping Physiological ADP-Ribosylation Using Activated Ion Electron Transfer Dissociation.

Buch-Larsen Sara C SC Hendriks Ivo A IA Lodge Jean M JM Rykær Martin M Furtwängler Benjamin B Shishkova Evgenia E Westphall Michael S MS Coon Joshua J JJ Nielsen Michael L ML

Cell reports 20200901 12

ADP-ribosylation (ADPr) is a post-translational modification that plays pivotal roles in a wide range of cellular processes. Mass spectrometry (MS)-based analysis of ADPr under physiological conditions, without relying on genetic or chemical perturbation, has been hindered by technical limitations. Here, we describe the applicability of activated ion electron transfer dissociation (AI-ETD) for MS-based proteomics analysis of physiological ADPr using our unbiased Af1521 enrichment strategy. To be ...[more]

PMID: 32966781

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