In:
Frontiers in Chemistry, Frontiers Media SA, Vol. 10 ( 2022-11-21)
Abstract:
Chemical cross-linking coupled with mass spectrometry has emerged as a powerful strategy which enables global profiling of protein interactome with direct interaction interfaces in complex biological systems. The alkyne-tagged enrichable cross-linkers are preferred to improve the coverage of low-abundance cross-linked peptides, combined with click chemistry for biotin conjugation to allow the cross-linked peptide enrichment. However, a systematic evaluation on the efficiency of click approaches (protein-based or peptide-based) and diverse cleavable click-chemistry ligands (acid, reduction, and photo) for cross-linked peptide enrichment and release is lacking. Herein, together with in vivo chemical cross-linking by alkyne-tagged cross-linkers, we explored the click-chemistry-based enrichment approaches on protein and peptide levels with three cleavable click-chemistry ligands, respectively. By comparison, the approach of protein-based click-chemistry conjugation with acid-cleavable tags was demonstrated to permit the most cross-linked peptide identification. The advancement of this strategy enhanced the proteome-wide cross-linking analysis, constructing a 5,518-protein–protein-interaction network among 1,871 proteins with widely abundant distribution in cells. Therefore, all these results demonstrated the guideline value of our work for efficient cross-linked peptide enrichment, thus facilitating the in-depth profiling of protein interactome for functional analysis.
Type of Medium:
Online Resource
ISSN:
2296-2646
DOI:
10.3389/fchem.2022.994572
DOI:
10.3389/fchem.2022.994572.s001
DOI:
10.3389/fchem.2022.994572.s002
DOI:
10.3389/fchem.2022.994572.s003
DOI:
10.3389/fchem.2022.994572.s004
DOI:
10.3389/fchem.2022.994572.s005
DOI:
10.3389/fchem.2022.994572.s006
DOI:
10.3389/fchem.2022.994572.s007
Language:
Unknown
Publisher:
Frontiers Media SA
Publication Date:
2022
detail.hit.zdb_id:
2711776-5
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