PXD076709 is an
original dataset announced via ProteomeXchange.
Dataset Summary
| Title | PARP14c Acidic Modification Hydroxylamine Optimization |
| Description | ADP-ribosylation is a highly dynamic process that is increasingly linked to a diverse set of cellular functions and pathophysiological outcomes. The modification of target substrates (e.g., proteins, DNA, RNA) involves ligation to an array of different chemical moieties. Recent work has shown that the ester bonds formed between ribose and Glu/Asp residues are particularly labile. A common strategy to overcome this lability is the replacement of the ADP-ribose (ADPr) by hydroxylamine. Attachment of hydroxamic acid (HA) permanently shifts the mass of labeled Glu/Asp residues by +15 Da. However, this method suffers from two notable limitations: HA treatment for long periods of time will label Glu/Asp sites that are solvent exposed, but not modified by ADPr; and the competing hydrolysis reaction still occurs in the conditions routinely used for HA labeling. Herein, we optimize a mass spectrometry (MS) workflow that overcomes these drawbacks. As a proof-of-principle, a truncated form of PARP14 was assessed for its ability to ADP-ribosylate peptides and itself. Conditions that prioritize HA attack over the background rate of hydrolysis were determined using an ultrathin matrix-assisted laser-desorption/ionization (TLC-MALDI) time-of-flight (TOF) method with ADPr-peptides. These efforts doubled HA labeling efficiency and completely removed ADPr within an hour. Next, a series of MS experiments identified the sites of PARP14 automodification using both standard HA labeling conditions (overnight, pH 7.0, 25 °C) and the optimized conditions. Of the identified 9 ADPr sites on PARP14, 5 are unique to the optimized conditions and 6 are newly identified. Mutagenesis revealed a set of sites that down-regulated PARP14 and another set that appears to activate PARP14. Taken together, these efforts have improved the true positive site identification rate using HA and have revealed a novel role for several automodification sites in PARP14 regulation. |
| HostingRepository | PRIDE |
| AnnounceDate | 2026-08-15 |
| AnnouncementXML | Submission_2026-08-14_16:54:36.100.xml |
| DigitalObjectIdentifier | https://doi.org/10.6019/PXD076709 |
| ReviewLevel | Peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Supported dataset by repository |
| PrimarySubmitter | Ian Carter-O'Connell |
| SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606; |
| ModificationList | Hydroxamic_acid; monohydroxylated residue; iodoacetamide derivatized residue |
| Instrument | Orbitrap Eclipse |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|
| 0 | 2026-04-07 13:52:29 | ID requested | |
| ⏵ 1 | 2026-08-14 16:54:36 | announced | |
Publication List
Keyword List
Contact List
| Ian Carter-O'Connell |
|---|
| contact affiliation | Department of Chemistry and Biochemistry, Santa Clara University, Santa Clara, CA 95053 USA |
| contact email | icarteroconnell@scu.edu |
| lab head | |
| Ian Carter-O'Connell |
|---|
| contact affiliation | Santa Clara University |
| contact email | icarteroconnell@scu.edu |
| dataset submitter | |
Full Dataset Link List
Dataset FTP location
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| PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD076709
- Label: PRIDE project
- Name: PARP14c Acidic Modification Hydroxylamine Optimization
