PXD026393
PXD026393 is an original dataset announced via ProteomeXchange.
Dataset Summary
| Title | Photoredox-catalyzed decarboxylative C-terminal differentiation for bulk and single molecule proteomics |
| Description | Methods for the selective labeling of functional groups on peptides are being developed and used in the workflow of both current and emerging proteomics technologies, such as single-molecule fluorosequencing. Photoredox methods in discriminative targeting of the terminal carboxylic acid from the internal acidic residues have been described but not effectively applied in concert with peptide-centric proteomic technologies. In this work, we describe methods for the use of lumiflavin based photoredox chemistry in peptide mass-spectrometry and single molecule protein sequencing. After optimizing the instrumentation and reaction conditions with the peptide angiotensin, we demonstrate successfully labeling of peptides in complex mixtures generated from enzymatic digests from bovine serum albumin, as well as protein mixtures from yeast and human cell extracts. Using two distinct analytical approaches, we characterize labeling efficiencies of greater than 50% across full proteomes and determine biases due to different C-terminal amino acids, finding histidine to be preferentially disfavored. Finally we demonstrate the ability to fluorescently label an internal acidic residue following C-terminal carboxylic acid derivatization, as suitable for single molecule protein sequencing technologies. The strategies and limitations described in this work provide clear guidance for further improvement and general use of this chemistry for analyzing proteins. |
| HostingRepository | MassIVE |
| AnnounceDate | 2021-12-10 |
| AnnouncementXML | Submission_2021-12-10_09:01:46.616.xml |
| DigitalObjectIdentifier | |
| ReviewLevel | Non peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Supported dataset by repository |
| PrimarySubmitter | Brendan Floyd |
| SpeciesList | scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606; scientific name: Saccharomyces cerevisiae; common name: baker's yeast; NCBI TaxID: 4932; scientific name: Bos taurus; common name: cattle; NCBI TaxID: 9913; |
| ModificationList | unknown modification |
| Instrument | Orbitrap Fusion |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|---|---|---|
| 0 | 2021-06-01 07:59:02 | ID requested | |
| ⏵ 1 | 2021-12-10 09:01:46 | announced |
Publication List
| no publication |
Keyword List
| submitter keyword: C-terminal, Bioconjugation |
Contact List
| Edward Marcotte | |
|---|---|
| contact affiliation | University of Texas-Austin |
| contact email | marcotte@icmb.utexas.edu |
| lab head | |
| Brendan Floyd | |
| contact affiliation | University of Texas at Austin |
| contact email | bmfloyd@utexas.edu |
| dataset submitter | |
Full Dataset Link List
| MassIVE dataset URI |
| Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://massive.ucsd.edu/MSV000087551/ |
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