title: Synthesis of Arylazide- and Diazirine-Containing CrAsH-EDT2 Photoaffinity Probes.
abstract: Two photo-crosslinking biarsenical (CrAsH-EDT2 )-modified probes were synthesized that are expected to be useful tools for tetracysteine-labeled proteins to facilitate the co-affinity purification of their DNA binding sequences and interacting proteins. In addition, improvements for the synthesis of CrAsH-EDT2 and N(1) -(4-azido-2-nitrophenyl)hexane-1,6-diamine are reported. Both photoprobes effectively entered HeLa cells (and the nucleus) and were dependent on the tetracysteine motif in recombinant DMRT1 (doublesex and Mab3-related transcription factor) to induce fluorescence, suggesting that their crosslinking abilities can be exploited for the identification of nucleic acids and proteins associated with a protein of interest.
authors.affiliation: Department of Medicinal Chemistry and Institute for Therapeutics Discovery and Development, University of Minnesota, Minneapolis, MN, USA.|Interdisciplinary Center for Male Contraceptive Research and Drug Development, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Shameem S
authors.initials: SS
authors.lastname: Syeda
authors.affiliation: Interdisciplinary Center for Male Contraceptive Research and Drug Development, University of Kansas Medical Center, Kansas City, KS, USA.|Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Daren
authors.initials: D
authors.lastname: Rice
authors.affiliation: Department of Medicinal Chemistry and Institute for Therapeutics Discovery and Development, University of Minnesota, Minneapolis, MN, USA.|Interdisciplinary Center for Male Contraceptive Research and Drug Development, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Derek J
authors.initials: DJ
authors.lastname: Hook
authors.affiliation: Interdisciplinary Center for Male Contraceptive Research and Drug Development, University of Kansas Medical Center, Kansas City, KS, USA.|Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Leslie L
authors.initials: LL
authors.lastname: Heckert
authors.affiliation: Department of Medicinal Chemistry and Institute for Therapeutics Discovery and Development, University of Minnesota, Minneapolis, MN, USA.|Interdisciplinary Center for Male Contraceptive Research and Drug Development, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Gunda I
authors.initials: GI
authors.lastname: Georg
mesh_terms: D001151:Arsenic; D001152:Arsenicals / Q000138:chemical synthesis / Q000737:chemistry*; D001386:Azides / Q000138:chemical synthesis / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D003959:Diamines / Q000138:chemical synthesis / Q000737:chemistry*; D003978:Diazomethane / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry*; D005452:Fluoresceins / Q000138:chemical synthesis / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D008623:Mercaptoethanol / Q000031:analogs & derivatives* / Q000737:chemistry; D019920:Photoaffinity Labels / Q000138:chemical synthesis / Q000737:chemistry*; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000378:metabolism
chemical_list: C000613298:4',5'-bis(1,2,3-dithioarsolan-2-yl)fluorescein-5-carboxylic acid; D001152:Arsenicals; D001386:Azides; D003432:Cross-Linking Reagents; C110547:DMRT1 protein; D003959:Diamines; D005452:Fluoresceins; C000613301:N(1)-(4-azido-2-nitrophenyl)hexane-1,6-diamine; D019920:Photoaffinity Labels; D014157:Transcription Factors; D008623:Mercaptoethanol; D003978:Diazomethane; C031854:1,2-ethanedithiol; D001151:Arsenic
keywords: Biarsenical probe; Fluorescence; Photoaffinity probes; Tetracysteine-tagged recombinant DMRT1