Proteomics

Dataset Information

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Rapid and high coverage profile of human cysteineome-Figure 6

ABSTRACT: Cysteines play important roles in many biological activities, including protein folding, aging, signaling and etc. To identify and quantitatively profile the oxidation and reactivity of cysteines, cysteine-reactive compounds have been applied to covalently label and enrich the cysteines. And with mass spectrometry (MS), thousands of cysteines can be profiled at the same time with decoration of functional groups. However, with all the technology and methods developed to characterize cysteines, current datasets have only uncovered about 5% of the total cysteinome. Here we show a comprehensive and high coverage profile of more than 30,000 cysteines. With incorporation of new techniques - single-pot, solid phase-enhanced sample-preparation (SP3) and High-field asymmetric waveform ion mobility spectrometry (FAIMS), we have successfully identified as many as 14,306 cystienes in a single-shot experiment. With cysteine profiles in different cell lines, proteases and fractionations, our results have successfully expanded the cysteine library to a great extent.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Tianyang Yan  

LAB HEAD: Keri M. Backus

PROVIDER: PXD023058 | Pride | 2021-02-03

REPOSITORIES: Pride

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2020-10-28-kb-70min_FAIMS_3cv_-35_-45_-55--OTIT-CF3_SP3_isoTOP-HD-3.raw Raw
2020-11-10-kb-70min_FAIMS_100micron_3cv_-35_-45_-55--OTIT_HD-1.raw Raw
2020-11-10-kb-70min_FAIMS_100micron_3cv_-35_-45_-55--OTIT_HD-2.raw Raw
2020-11-10-kb-70min_FAIMS_100micron_3cv_-35_-45_-55--OTIT_HD-3.raw Raw
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Publications

SP3-FAIMS Chemoproteomics for High-Coverage Profiling of the Human Cysteinome*.

Yan Tianyang T   Desai Heta S HS   Boatner Lisa M LM   Yen Stephanie L SL   Cao Jian J   Palafox Maria F MF   Jami-Alahmadi Yasaman Y   Backus Keriann M KM  

Chembiochem : a European journal of chemical biology 20210218 10

Chemoproteomics has enabled the rapid and proteome-wide discovery of functional, redox-sensitive, and ligandable cysteine residues. Despite widespread adoption and considerable advances in both sample-preparation workflows and MS instrumentation, chemoproteomics experiments still typically only identify a small fraction of all cysteines encoded by the human genome. Here, we develop an optimized sample-preparation workflow that combines enhanced peptide labeling with single-pot, solid-phase-enhan  ...[more]

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