Mapping Physiological ADP-Ribosylation Using Activated Ion Electron Transfer Dissociation
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Mapping Physiological ADP-Ribosylation Using Activated Ion Electron Transfer Dissociation. / Buch-Larsen, Sara C.; Hendriks, Ivo A.; Lodge, Jean M.; Rykaer, Martin; Furtwangler, Benjamin; Shishkova, Evgenia; Westphall, Michael S.; Coon, Joshua J.; Nielsen, Michael L.
In: Cell Reports, Vol. 32, No. 12, 108176, 2020.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Mapping Physiological ADP-Ribosylation Using Activated Ion Electron Transfer Dissociation
AU - Buch-Larsen, Sara C.
AU - Hendriks, Ivo A.
AU - Lodge, Jean M.
AU - Rykaer, Martin
AU - Furtwangler, Benjamin
AU - Shishkova, Evgenia
AU - Westphall, Michael S.
AU - Coon, Joshua J.
AU - Nielsen, Michael L.
PY - 2020
Y1 - 2020
N2 - 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.
AB - 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.
KW - POLY(ADP-RIBOSYL)ATION
KW - PARP-1
KW - SITE
KW - IDENTIFICATION
KW - PROTEIN
KW - ASSOCIATION
KW - GENERATION
KW - MECHANISM
KW - ARGININE
KW - PRODUCT
U2 - 10.1016/j.celrep.2020.108176
DO - 10.1016/j.celrep.2020.108176
M3 - Journal article
C2 - 32966781
VL - 32
JO - Cell Reports
JF - Cell Reports
SN - 2211-1247
IS - 12
M1 - 108176
ER -
ID: 249864305