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Synthesis of Arylazide‐ and Diazirine‐Containing CrAsH‐EDT 2 Photoaffinity Probes
Author(s) -
Syeda Shameem S.,
Rice Daren,
Hook Derek J.,
Heckert Leslie L.,
Georg Gunda I.
Publication year - 2016
Publication title -
archiv der pharmazie
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 61
eISSN - 1521-4184
pISSN - 0365-6233
DOI - 10.1002/ardp.201500440
Subject(s) - diazirine , chemistry , nucleic acid , intein , dna , covalent bond , nucleus , recombinant dna , combinatorial chemistry , hela , transcription factor , dna binding protein , biochemistry , fluorescence , stereochemistry , biophysics , biology , microbiology and biotechnology , gene , in vitro , organic chemistry , rna , rna splicing , physics , quantum mechanics
Two photo‐crosslinking biarsenical (CrAsH‐EDT 2 )‐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‐EDT 2 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.