title: Divide and conquer: cleavable cross-linkers to study protein conformation and protein-protein interactions.
abstract: Chemical cross-linking combined with mass spectrometry (MS) and computational modeling has evolved as an alternative method to address fundamental questions in structural biology. The constraints revealed by the cross-links yield valuable distance information and allow one to deduce three-dimensional structural information on very large and transient protein complexes. During the past few years, technical advances in the cross-linking/MS approach have been enormous, mainly owing to the fantastic advances in MS technology, and it is easily overlooked that significant progress has been made in the design of novel cross-linking reagents. In this review, the advent of cleavable cross-linking reagents will be highlighted. In particular, gas-phase (MS-) cleavable cross-linkers offer unique properties for an automated, data-dependent assignment of cross-linked products based on the generation of characteristic fragment ion signatures in MS/MS and MS3spectra. Therefore, MS-cleavable cross-linkers are envisioned to hold the key for proteome-wide applications of the chemical cross-linking/MS approach, not only to delineate the conformation of single proteins but also to decipher protein interaction networks. Graphical Abstract Outline of the analytical strategy using cleavable cross-linkers in combination with MS to conduct protein conformational and protein interaction studies.
authors.affiliation: Department of Pharmaceutical Chemistry & Bioanalytics, Institute of Pharmacy, Martin-Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Str. 4, 06120, Halle (Saale), Germany. andrea.sinz@pharmazie.uni-halle.de.
authors.forename: Andrea
authors.initials: A
authors.lastname: Sinz
mesh_terms: D000818:Animals; D003432:Cross-Linking Reagents / Q000737:chemistry*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008958:Models, Molecular; D010084:Oxidation-Reduction; D011487:Protein Conformation; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D003432:Cross-Linking Reagents; D011506:Proteins
keywords: Chemical cross-linking; Cleavable cross-linkers; Mass spectrometry; Protein 3D structure; Protein interaction networks