title: A simple and effective cleavable linker for chemical proteomics applications.
abstract: The study of metabolically labeled or probe-modified proteins is an important area in chemical proteomics. Isolation and purification of the protein targets is a necessary step before MS identification. The biotin-streptavidin system is widely used in this process, but the harsh denaturing conditions also release natively biotinylated proteins and non-selectively bound proteins. A cleavable linker strategy is a promising approach for solving this problem. Though several cleavable linkers have been developed and tested, an efficient, easily synthesized, and inexpensive cleavable linker is a desirable addition to the proteomics toolbox. Here, we describe the chemical proteomics application of a vicinal diol cleavable linker. Through easy-to-handle chemistry we incorporate this linker into an activity-based probe and a biotin alkyne tag amenable for bioorthogonal ligation. With these reagents, background protein identifications are significantly reduced relative to standard on-bead digestion.
authors.affiliation: Lehrstuhl für Chemie der Biopolymere, Technische Universität München, Weihenstephaner Berg 3, 85354 Freising, Germany.
authors.forename: Yinliang
authors.initials: Y
authors.lastname: Yang
authors.forename: Hannes
authors.initials: H
authors.lastname: Hahne
authors.forename: Bernhard
authors.initials: B
authors.lastname: Kuster
authors.forename: Steven H L
authors.initials: SH
authors.lastname: Verhelst
mesh_terms: D001710:Biotin / Q000737:chemistry; D002403:Cathepsins; D003432:Cross-Linking Reagents / Q000737:chemistry*; D005960:Glucosides / Q000737:chemistry; D013058:Mass Spectrometry; D011506:Proteins; D020543:Proteome / Q000032:analysis / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D011744:Pyrimidinones / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D005960:Glucosides; D011506:Proteins; D020543:Proteome; D011744:Pyrimidinones; D001710:Biotin; C009661:vicine; D002403:Cathepsins