title: A universal matrix-assisted laser desorption/ionization cleavable cross-linker for protein structure analysis.
abstract: The concept of protein cross-linking in combination with mass spectrometry holds great promise to derive structural information on protein conformation and protein-protein interactions. We recently presented a dissociative amine-reactive cross-linker (NHS-BuUrBu-NHS) that is shown herein to be universally applicable to protein structure analysis under matrix-assisted laser desorption/ionization tandem mass spectrometric (MALDI-MS/MS) conditions, based on the examples of the peptides substance P, luteinizing hormone releasing hormone (LHRH), and the 32-kDa ligand-binding domain of peroxisome proliferator-activated receptor alpha (PPARα). The characteristic fragment ion patterns and constant neutral losses of the cross-linker greatly simplify the identification of different cross-linked species from complex mixtures and drastically reduce the potential of identifying false-positive cross-links. Therefore, this cross-linker holds an enormous potential for deriving structural information of proteins and protein complexes in a highly automated fashion.
authors.affiliation: Department of Pharmaceutical Chemistry & Bioanalytics, Institute of Pharmacy, Martin-Luther University Halle-Wittenberg, D-06120 Halle (Saale), Germany.
authors.forename: Mathias Q
authors.initials: MQ
authors.lastname: Müller
authors.forename: Johannes J
authors.initials: JJ
authors.lastname: Zeiser
authors.forename: Frank
authors.initials: F
authors.lastname: Dreiocker
authors.forename: Andreas
authors.initials: A
authors.lastname: Pich
authors.forename: Mathias
authors.initials: M
authors.lastname: Schäfer
authors.forename: Andrea
authors.initials: A
authors.lastname: Sinz
mesh_terms: D020148:Butyric Acid / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013388:Succinimides / Q000737:chemistry; D014508:Urea / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D011506:Proteins; D013388:Succinimides; D020148:Butyric Acid; C035761:N-hydroxysulfosuccinimide; D014508:Urea