Design and Application of Novel Cross-Linking Reagents for Mapping Protein-Protein Interactions

摘要

Characterization of dynamic protein interactions is a challenging endeavor because a single protein may interact transiently with various other molecules within the cell. Chemical cross-linking coupled with tandem mass spectrometry has been successfully employed to overcome the challenges of studying dynamic protein complexes. However, this potentially powerful strategy is often complicated by the presence of mixtures of cross-linked and unmodified proteins. To enhance our ability to detect cross-linked peptides and elucidate protein structure, we have designed and synthesized a novel cross-linker which is effective for enrichment and subsequent analysis. We have explored both dipolar cycloaddition with a biotinylated alkyne and Staudinger ligation with a biotinylated phosphine for enrichment.