MassIVE MSV000095995

Partial Public PXD056407

Establishing a Top-Down Proteomics Platform on a Time-of-Flight Instrument with Electron-Activated Dissociation

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Description

. Top-down proteomics is the study of intact proteins and their post-translational modifications with mass spectrometry. Historically, this field is more challenging than its bottom-up counterpart because the species are much bigger and have a larger number of possible combinations of sequences and modifications, thus there is a great need for technological development. With improvements in instrumentation and a multiplicity of fragmentation modes available, top-down proteomics is quickly gaining in popularity with renewed attention on increasing confidence in identification and quantification. Here, we systematically evaluated the Sciex ZenoTOF 7600 system for top-down proteomics, applying standards in the field to validate the platform and further experimenting with its capabilities in electron-activated dissociation and post-translational modification site localization. The instrument demonstrated robustness in standard proteins for platform QC, as aided by zeno-trapping. We were also able to apply this to histone post-translational modifications, achieving high sequence coverage that allowed PTMs site-localization across protein sequences with optimized EAD fragmentation. We demonstrated the ability to analyze proteins spanning the mass range and included analysis of glycosylated proteins. This is a reference point for future top-down proteomics experiments to be conducted on the ZenoTOF 7600 system. [doi:10.25345/C51R6NC1Z] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Top-Down Proteomics

Contact

Principal Investigators:
(in alphabetical order) 		Benjamin A. Garcia, Department of Biochemistry and Biophysics, University of Pennsylvania, N/A
Submitting User: rsearfoss
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Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

"N/A" means no results of this type were submitted.
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