Development of a compact alkynyl-enrichable crosslinker for in-depth in-vivo crosslinking analysis
Gao, H.; Zhao, L. L.; Zhao, Q.; Zhang, H. L.; Zhao, F. B.; Gong, Z.
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Chemical crosslinking coupled with mass spectrometry (CXMS) has emerged as a powerfl technique to capture the dynamic information of protein complexes with high sensitivity, throughput and sample universality. To advance the study of in-vivo protein structures and protein-protein interactions on the large scale, a new alkynyl-enrichable crosslinker was developed with high efficiency of membrane penetration, reactivity and enrichment. The crosslinker was successfully used for in-vivo crosslinking of intact human cells, reslting in 6820 non-redundant crosslinks identified at a false discovery rate (FDR) of 1% using pLink 2.0, which 4898 (71.8%) of the cross-links were assigned as intraprotein and 1922 (28.2%) were interprotein links. To our knowledge, this is also the first time to realize the in-vivo crosslinking with a non-cleavable crosslinker for homo species cells.
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