title: 2-Aryl-5-carboxytetrazole as a New Photoaffinity Label for Drug Target Identification.
abstract: Photoaffinity labels are powerful tools for dissecting ligand-protein interactions, and they have a broad utility in medicinal chemistry and drug discovery. Traditional photoaffinity labels work through nonspecific C-H/X-H bond insertion reactions with the protein of interest by the highly reactive photogenerated intermediate. Herein, we report a new photoaffinity label, 2-aryl-5-carboxytetrazole (ACT), that interacts with the target protein via a unique mechanism in which the photogenerated carboxynitrile imine reacts with a proximal nucleophile near the target active site. In two distinct case studies, we demonstrate that the attachment of ACT to a ligand does not significantly alter the binding affinity and specificity of the parent drug. Compared with diazirine and benzophenone, two commonly used photoaffinity labels, in two case studies ACT showed higher photo-cross-linking yields toward their protein targets in vitro based on mass spectrometry analysis. In the in situ target identification studies, ACT successfully captured the desired targets with an efficiency comparable to the diazirine. We expect that further development of this class of photoaffinity labels will lead to a broad range of applications across target identification, and validation and elucidation of the binding site in drug discovery.
authors.affiliation: Department of Chemistry, State University of New York at Buffalo , Buffalo, New York 14260-3000, United States.
authors.forename: András
authors.initials: A
authors.lastname: Herner
authors.affiliation: Discovery Chemistry and Technology, AbbVie Inc. , North Chicago, Illinois 60064-6101, United States.
authors.forename: Jasmina
authors.initials: J
authors.lastname: Marjanovic
authors.affiliation: Department of Chemistry, State University of New York at Buffalo , Buffalo, New York 14260-3000, United States.
authors.forename: Tracey M
authors.initials: TM
authors.lastname: Lewandowski
authors.affiliation: Discovery Chemistry and Technology, AbbVie Inc. , North Chicago, Illinois 60064-6101, United States.
authors.forename: Violeta
authors.initials: V
authors.lastname: Marin
authors.affiliation: Discovery Chemistry and Technology, AbbVie Inc. , North Chicago, Illinois 60064-6101, United States.
authors.forename: Melanie
authors.initials: M
authors.lastname: Patterson
authors.affiliation: Discovery Chemistry and Technology, AbbVie Inc. , North Chicago, Illinois 60064-6101, United States.
authors.forename: Laura
authors.initials: L
authors.lastname: Miesbauer
authors.affiliation: Discovery Chemistry and Technology, AbbVie Inc. , North Chicago, Illinois 60064-6101, United States.
authors.forename: Damien
authors.initials: D
authors.lastname: Ready
authors.affiliation: Discovery Chemistry and Technology, AbbVie Inc. , North Chicago, Illinois 60064-6101, United States.
authors.forename: Jon
authors.initials: J
authors.lastname: Williams
authors.affiliation: Discovery Chemistry and Technology, AbbVie Inc. , North Chicago, Illinois 60064-6101, United States.
authors.forename: Anil
authors.initials: A
authors.lastname: Vasudevan
authors.affiliation: Department of Chemistry, State University of New York at Buffalo , Buffalo, New York 14260-3000, United States.
authors.forename: Qing
authors.initials: Q
authors.lastname: Lin
mesh_terms: D000595:Amino Acid Sequence; D019295:Computational Biology; D000069439:Dasatinib / Q000737:chemistry; D016503:Drug Delivery Systems; D005453:Fluorescence; D006801:Humans; D020014:K562 Cells; D015394:Molecular Structure; D019920:Photoaffinity Labels / Q000737:chemistry*; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D013777:Tetrazoles / Q000737:chemistry*
chemical_list: D019920:Photoaffinity Labels; D013777:Tetrazoles; D000069439:Dasatinib