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Carboxylate-Selective Chemical Cross-Linkers for Mass Spectrometric Analysis of Protein Structures.

Xiaoyun ZhangJian-Hua WangDan TanQiang LiMaodong LiZhou GongChun TangZhirong LiuMeng-Qiu DongXiaoguang Lei
Published in: Analytical chemistry (2018)
Chemical cross-linking coupled with mass spectrometry (CXMS) facilitates structural analysis of proteins. As current CXMS applications are almost exclusively limited to lysine residues, they can only retrieve a small portion of the structural information theoretically accessible to CXMS. Chemical cross-linkers targeting the acidic residues Asp/Glu could greatly enhance the power of CXMS. However, it has been difficult to develop chemistries that offer selectivity and efficiency under physiological conditions. Here, we report a class of carboxylate-selective diazo-containing cross-linkers (Diazoker) of which Diazoker 1, with a spacer arm consisting of two ethan-1,2-diol units, is the best example. Unlike previously developed carboxylate-selective cross-linkers like pimelic acid dihydrazide (PDH), Diazoker 1 does not require a coupling reagent. We tested Diazoker 1 on nine model proteins and found that Diazoker 1 generated an average of 73 cross-linked peptide pairs per protein. Although this is 32% fewer than the number generated by PDH, the Diazoker 1 cross-links have a higher rate of compatibility with protein crystal structures. From a more complex protein mixture, Diazoker 1 and PDH identified 75 and 76 cross-linked peptide pairs, respectively. The Asp/Glu residues cross-linked by Diazoker 1 are not the same as those cross-linked by PDH. Diazoker 1 favors acidic residues that are less exposed to solvent. In conclusion, Diazoker 1 is complementary to existing cross-linkers and expands the toolkit of CXMS for structural analysis of proteins.
Keyphrases
  • mass spectrometry
  • amino acid
  • protein protein
  • ionic liquid
  • binding protein
  • healthcare
  • high resolution
  • drug delivery
  • cancer therapy
  • electron transfer