title: Rational design of a photo-crosslinking BODIPY for in situ protein labeling.
abstract: Photo-crosslinking agents have emerged as critical tools to investigate protein-protein interactions in complex proteomes, but there are few photocrosslinkers available at the moment. Here, we report the first rational design of a photo-crosslinking BODIPY fluorophore (pcBD) and its biological application for biomolecule labeling. As a photosensitizing functional motif, an aryl ketone group was incorporated into the BODIPY fluorophore, and a series of proteins were labeled by pcBD compounds upon UV irradiation. In order to investigate protein-protein interactions in a protein mixture, amino-functionalized pcBD was prepared and covalently attached to a ubiquitin ligase binding peptide. Upon UV irradiation, we could successfully visualize the substrates in the total lysate. These results provided a proof of concept for spatially controllable tagging via photo-activation of the pcBD scaffold, and demonstrated its potential usage for in situ labeling applications.
authors.affiliation: Molecular Recognition Research Center, Korea Institute of Science and Technology, 39-1 Hawolgok-dong, Seoul, 136-791, Republic of Korea. junseoklee@kist.re.kr.
authors.forename: Dhiraj P
authors.initials: DP
authors.lastname: Murale
authors.forename: Seong Cheol
authors.initials: SC
authors.lastname: Hong
authors.forename: JiHyeon
authors.initials: J
authors.lastname: Yun
authors.forename: Chang No
authors.initials: CN
authors.lastname: Yoon
authors.forename: Jun-Seok
authors.initials: JS
authors.lastname: Lee
mesh_terms: D001896:Boron Compounds / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D015195:Drug Design; D005456:Fluorescent Dyes / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D055668:Photochemical Processes; D013194:Staining and Labeling; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry*
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D044767:Ubiquitin-Protein Ligases