MassIVE MSV000092853

Partial Public PXD045359

A systematic and validated Fe(II)-EDTA method of hydroxyl radical generation for oxidative protein footprinting

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Description

Correlating the structure and dynamics of proteins with biological function is critical to understanding normal and dysfunctional cellular mechanisms. We describe a quantitative method of hydroxyl radical generation via Fe(II)-EDTA catalyzed Fenton chemistry that provides easy access to oxidative protein footprinting using equipment commonly found in research and process control laboratories. Robust and reproducible dose-dependent oxidation of protein samples is observed and quantitated by mass spectrometry with as fine as single residue resolution. An oxidation analysis of lysozyme provides a readily accessible benchmark for our method. The efficacy of our oxidation method is demonstrated through mapping the interface of a RAS monobody complex, the surface of the NIST mAb, and the interface between PRC2 complex components. These studies are executed using standard laboratory tools and a few pennies of reagents; the mass spectrometry analysis can be streamlined to map protein structure with single amino acid residue resolution. All mass spectrometry raw files for this study are included here. [doi:10.25345/C57H1DX7P] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Fenton reaction ; oxidative footprinting

Contact

Principal Investigators:
(in alphabetical order) 		Beatrix Ueberheide, NYU Langone Health, USA
Submitting User: Trixi
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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.

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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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