Synthesis of Arylazide- and Diazirine-Containing CrAsH-EDT2 Photoaffinity Probes

• Published in: Archiv der Pharmazie (Weinheim) Vol. 349; no. 4; pp. 233 - 241
• Main Authors: Syeda, Shameem S., Rice, Daren, Hook, Derek J., Heckert, Leslie L., Georg, Gunda I.
• Format: Journal Article
• Language: English; German
• Published: WEINHEIM Blackwell Publishing Ltd 01.04.2016; Wiley; Wiley Subscription Services, Inc; John Wiley and Sons Inc
• Subjects: Arsenic; Arsenicals - chemical synthesis; Arsenicals - chemistry; Azides - chemical synthesis; Azides - chemistry; Biarsenical probe; Chemistry; Chemistry, Medicinal; Chemistry, Multidisciplinary; Cross-Linking Reagents - chemistry; Diamines - chemical synthesis; Diamines - chemistry; Diazomethane - analogs & derivatives; Diazomethane - chemical synthesis; Diazomethane - chemistry; Fluoresceins - chemical synthesis; Fluoresceins - chemistry; Fluorescence; HeLa Cells; Humans; Life Sciences & Biomedicine; Mercaptoethanol - analogs & derivatives; Mercaptoethanol - chemistry; Pharmacology & Pharmacy; Photoaffinity Labels - chemical synthesis; Photoaffinity Labels - chemistry; Photoaffinity probes; Physical Sciences; Proteins; Science & Technology; Structure-Activity Relationship; Tetracysteine-tagged recombinant DMRT1; Transcription Factors - metabolism; CELLS; Photoaffinity probes; TETRACYSTEINE-TAGGED PROTEINS; Fluorescence; IDENTIFICATION; AFFINITY PROBE; PHOTO-CROSS-LINKING; Tetracysteine-tagged recombinant DMRT1; IN-VIVO; Biarsenical probe; BENZOPHENONE; CHEMISTRY; BINDING
• ISSN: 0365-6233; 1521-4184
• DOI: 10.1002/ardp.201500440

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 N1‐(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. Two photo‐crosslinking biarsenical probes were synthesized that are expected to be useful tools for identifying the DNA binding sequences and interacting proteins of tetracysteine‐labeled proteins, by co‐affinity purification. Both photoprobes effectively entered HeLa cells (and the nucleus) and were dependent on the tetracysteine motif in recombinant DMRT1 to induce fluorescence.