title: Genetically encoded cleavable protein photo-cross-linker.
abstract: We have developed a genetically encoded, selenium-based cleavable photo-cross-linker that allows for the separation of bait and prey proteins after protein photo-cross-linking. We have further demonstrated the efficient capture of the in situ generated selenenic acid on the cleaved prey proteins. Our strategy involves tagging the selenenic acid with an alkyne-containing dimethoxyaniline molecule and subsequently labeling with an azide-bearing fluorophore or biotin probe. This cleavage-and-capture after protein photo-cross-linking strategy allows for the efficient capture of prey proteins that are readily accessible by two-dimensional gel-based proteomics and mass spectrometry analysis.
authors.affiliation: Beijing National Laboratory for Molecular Sciences, Synthetic and functional Biomolecules Center, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry, and Molecular Engineering, Peking University , Beijing 100871, China.
authors.forename: Shixian
authors.initials: S
authors.lastname: Lin
authors.forename: Dan
authors.initials: D
authors.lastname: He
authors.forename: Teng
authors.initials: T
authors.lastname: Long
authors.forename: Shuai
authors.initials: S
authors.lastname: Zhang
authors.forename: Rong
authors.initials: R
authors.lastname: Meng
authors.forename: Peng R
authors.initials: PR
authors.lastname: Chen
mesh_terms: D001665:Binding Sites; D001710:Biotin / Q000737:chemistry / Q000235:genetics; D002853:Chromatography, Liquid; D003432:Cross-Linking Reagents / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics; D006861:Hydrogen Peroxide / Q000737:chemistry; D008956:Models, Chemical; D016566:Organoselenium Compounds / Q000138:chemical synthesis / Q000737:chemistry*; D055668:Photochemical Processes; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000737:chemistry / Q000235:genetics; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D003432:Cross-Linking Reagents; D016566:Organoselenium Compounds; D011506:Proteins; D049452:Green Fluorescent Proteins; D001710:Biotin; D006861:Hydrogen Peroxide