Sequential digestion with Trypsin and Elastase in cross-linking/mass spectrometry

ABSTRACT: Cross-linking/mass spectrometry has become an important approach for studying protein structures and protein-protein interactions. The amino acid composition of some protein regions impedes the detection of cross-linked residues, although it would yield invaluable information for protein modelling. Here, we report on a sequential digestion strategy with trypsin and elastase to penetrate regions with a low density of trypsin cleavage sites. We exploited intrinsic substrate recognition proper-ties of elastase to specifically target larger tryptic peptides. Our application of this protocol to the TAF4-12 complex allowed us to identify cross-links in previously inaccessible regions.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human) Schizosaccharomyces Pombe

SUBMITTER: Therese Dau  

LAB HEAD: Juri Rappsilber

PROVIDER: PXD011459 | Pride | 2019-03-07

REPOSITORIES: Pride