title: A new crosslinker for mass spectrometric analysis of the quaternary structure of protein complexes.
abstract: Mass spectrometric structural analysis of crosslinked peptides is a powerful method to elucidate the spatial arrangement of polypeptides in protein complexes. Our aim is to develop bifunctional crosslinkers that, after crosslinking protein complexes followed by proteolytic digestion, give rise to crosslinked peptides that can be readily tracked down by mass spectrometry. To this end we synthesized the crosslinker N-benzyliminodiacetoyloxysuccinimid (BID), which yields stable benzyl cation marker ions upon low-energy collision-induced dissociation (CID) tandem mass spectrometry. Sensitive detection of the marker ion upon low-energy CID is demonstrated with different BID-crosslinked peptide preparations. With BID it becomes possible to retrieve crosslinked and crosslinker-adducted peptides, without the necessity of purifying crosslinked peptides prior to identification. The basic design of this crosslinker can be varied upon, in order to meet specific crosslinking needs.
authors.affiliation: Mass Spectrometry Group, Swammerdam Institute for Life Sciences, University of Amsterdam, The Netherlands.
authors.forename: J W
authors.initials: JW
authors.lastname: Back
authors.forename: A F
authors.initials: AF
authors.lastname: Hartog
authors.forename: H L
authors.initials: HL
authors.lastname: Dekker
authors.forename: A O
authors.initials: AO
authors.lastname: Muijsers
authors.forename: L J
authors.initials: LJ
authors.lastname: de Koning
authors.forename: L
authors.initials: L
authors.lastname: de Jong
mesh_terms: D001920:Bradykinin / Q000031:analogs & derivatives / Q000737:chemistry; D003432:Cross-Linking Reagents; D007098:Imino Acids; D009682:Magnetic Resonance Spectroscopy; D009496:Neurotensin / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D003432:Cross-Linking Reagents; D007098:Imino Acids; C421865:N-benzyliminodiacetoyloxysuccinimid; D011506:Proteins; C421866:Tyr-Lys-bradykinin; D009496:Neurotensin; D001920:Bradykinin