Chang Ho Sohn, Heather D Agnew, J Eugene Lee, Michael J Sweredoski, Robert L J Graham, Geoffrey T Smith, Sonja Hess, Gregg Czerwieniec, Joseph A Loo, James R Heath, Raymond J Deshaies, J L Beauchamp
We present novel homobifunctional amine-reactive clickable cross-linkers (CXLs) for investigation of three-dimensional protein structures and protein-protein interactions (PPIs). CXLs afford consolidated advantages not previously available in a simple cross-linker, including (1) their small size and cationic nature at physiological pH, resulting in good water solubility and cell-permeability, (2) an alkyne group for bio-orthogonal conjugation to affinity tags via the click reaction for enrichment of cross-linked peptides, (3) a nucleophilic displacement reaction involving the 1,2,3-triazole ring formed in the click reaction, yielding a lock-mass reporter ion for only clicked peptides, and (4) higher charge states of cross-linked peptides in the gas-phase for augmented electron transfer dissociation (ETD) yields...
March 20, 2012: Analytical Chemistry