AI Article Synopsis
- Cross-linking mass spectrometry (XL-MS) is a method for analyzing protein structures and interactions but struggles with in vivo studies due to inefficiencies in cross-linking reagents.
- Researchers developed a new cross-linker, tBu-PhoX, that can effectively penetrate cell membranes and allows for efficient enrichment of cross-linked peptides.
- This new approach significantly increases the number of identifiable cross-links in living human cells while reducing the time required for analysis, advancing the study of protein interactions in living systems.
Article Abstract
Cross-linking mass spectrometry (XL-MS) is an attractive method for the proteome-wide characterization of protein structures and interactions. Currently, the depth of in vivo XL-MS studies is lagging behind the established applications to cell lysates, because cross-linking reagents that can penetrate intact cells and strategies to enrich cross-linked peptides lack efficiency. To tackle these limitations, we have developed a phosphonate-containing cross-linker, tBu-PhoX, that efficiently permeates various biological membranes and can be robustly enriched using routine immobilized metal ion affinity chromatography. We have established a tBu-PhoX-based in vivo XL-MS approach that enables cross-links in intact human cells to be identified in high numbers with substantially reduced analysis time. Collectively, the developed cross-linker and XL-MS approach pave the way for the comprehensive XL-MS characterization of living systems.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9303249 | PMC |
| http://dx.doi.org/10.1002/anie.202113937 | DOI Listing |
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