Chemical cross-linking/mass spectrometry targeting acidic residues in proteins and protein complexes.

ABSTRACT: The study of proteins and protein complexes using chemical cross-linking followed by the MS identification of the cross-linked peptides has found increasingly widespread use in recent years. Thus far, such analyses have used almost exclusively homobifunctional, amine-reactive cross-linking reagents. Here we report the development and application of an orthogonal cross-linking chemistry specific for carboxyl groups. Chemical cross-linking of acidic residues is achieved using homobifunctional dihydrazides as cross-linking reagents and a coupling chemistry at neutral pH that is compatible with the structural integrity of most protein complexes. In addition to cross-links formed through insertion of the dihydrazides with different spacer lengths, zero-length cross-link products are also obtain

SUBMITTER: Leitner A 

PROVIDER: S-EPMC4084482 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature