Proteomics

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Ammonium Sulfate-based Prefractionation Improved Carbonylated Proteome Coverage in Arabidopsis thaliana Leaf Extract

ABSTRACT: The hypothesis that ammonium sulfate-based protein precipitation can help prefractionate the total protein extract into fractions of reduced complexity and improve the detection of low-abundance proteins was investigated. This dataset resulted from the experiments that we performed to improve the detection of the carbonylated proteins in leaf extract. A total of 122 carbonylated proteins were identified, of which 77 were found in the ammonium sulfate fractions only. This prefractionation method allowed to identify 63% more carbonylated proteins compared to the number of carbonylated proteins identified from the total crude extract without prefractionation. Our results thus showed that a simple offline prefractionation of the proteome by ammonium sulfate prior to liquid chromatography-tandem mass spectrometry significantly improved the detection of carbonylated proteins in Arabidopsis leaf extract.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Arabidopsis Thaliana (ncbitaxon:3702)

SUBMITTER: Tagnon Missihoun  

PROVIDER: MSV000090559 | MassIVE | Thu Oct 20 11:25:00 BST 2022

SECONDARY ACCESSION(S): PXD037611

REPOSITORIES: MassIVE

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Ammonium sulfate-based prefractionation improved proteome coverage and detection of carbonylated proteins in Arabidopsis thaliana leaf extract.

Tola Adesola Julius AJ   Missihoun Tagnon D TD  

Planta 20230219 3

<h4>Main conclusion</h4>Ammonium sulfate is well known to salt out proteins at high concentrations. The study revealed that it can serve to increase by 60% the total number of identified carbonylated proteins by LC-MS/MS. Protein carbonylation is a significant post-translational modification associated with reactive oxygen species signaling in animal and plant cells. However, the detection of carbonylated proteins involved in signaling is still challenging, as they only represent a small subset  ...[more]

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