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Quantitative analysis of in vivo methionine oxidation of the human proteome

ABSTRACT: A novel stable isotope labelling strategy was developed to quantify methionine oxidation in an unstressed human proteome. Cell extracts were oxidized with 18O labelled hydrogen peroxide following cell lysis in order to convert all unoxidized methionine residues to an oxidized version with a heavy label. The heavy labelled peptides were used to generate a custom search and quantification of the relative ratios between heavy and light labelled methionine sulfoxide containing peptides. Where the light labelled peptides were in vivo oxidized

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: John Bettinger

LAB HEAD: Sina Ghaemmaghami

PROVIDER: PXD014629 | Pride | 2019-12-09

REPOSITORIES: Pride

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Quantitative Analysis of in Vivo Methionine Oxidation of the Human Proteome.

Bettinger John Q JQ Welle Kevin A KA Hryhorenko Jennifer R JR Ghaemmaghami Sina S

Journal of proteome research 20200107 2

The oxidation of methionine is an important post-translational modification of proteins with numerous roles in physiology and pathology. However, the quantitative analysis of methionine oxidation on a proteome-wide scale has been hampered by technical limitations. Methionine is readily oxidized in vitro during sample preparation and analysis. In addition, there is a lack of enrichment protocols for peptides that contain an oxidized methionine residue, making the accurate quantification of methio ...[more]

PMID: 31801345

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Quantitative analysis of in vivo methionine oxidation of the human proteome

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Accurate proteome-wide measurement of methionine oxidation in aging mouse brains

Project description:The oxidation of methionine side chains has emerged as an important posttranslational modification of proteins. A diverse array of low-throughput and targeted studies have suggested that the oxidation of methionine residues in select proteins can have diverse impacts on cell physiology, ranging from detrimental effects on protein structure and stability to functional roles in cell signaling. Despite its importance, the large-scale investigation of methionine oxidation in a complex matrix, such as the cellular proteome has been historically hampered by technical limitations. Herein we report a methodology, Methionine Oxidation by Blocking (MobB), that allows for accurate and precise quantification of low levels of methionine oxidation typically observed in vivo. To demonstrate the utility of this methodology, we applied MobB to the brain tissues of young (6 m.o.) and old (20 m.o.) mice and identified over 280 novel sites for in vivo methionine oxidation. We further demonstrated that oxidation stoichiometries for specific methionine residues are highly consistent between individual animals and methionine sulfoxides are enriched in clusters of functionally related gene products including membrane and extracellular proteins. However, we did not detect significant changes in methionine oxidation in brains of old mice, suggesting that oxidation levels are highly regulated during the course of aging.

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Identification of PM-localized proteins

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c-Abl Dependent Tyrosine Phosphorylation of theCo-chaperone Aha1 Regulates Interaction with Hsp90

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The Quantitative Changes in the Yeast Hsp70 and Hsp90 Interactomes upon DNA Damage

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A hybrid structural determination approach to investigate protein-RNA complexes of the nucleocapsid protein of SARS-CoV-2

Project description:UV-light-induced protein-RNA cross-linking followed by MS analysis was used to identify the interaction sites between the N-terminal (NTD) or C-terminal domain (CTD) of the SARS-CoV-2 nucleocapsid protein and the s2m element of the viral RNA.

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N-glycosylation site occupancy in human prostaglandin H synthases expressed in Pichia pastoris

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