Syeda Shameem S, Rice Daren, Hook Derek J, Heckert Leslie L, Georg Gunda I
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.
Arch Pharm (Weinheim). 2016 Apr;349(4):233-41. doi: 10.1002/ardp.201500440. Epub 2016 Mar 7.
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.
合成了两种光交联双砷（CrAsH-EDT2）修饰的探针，预计它们将成为用于四半胱氨酸标记蛋白质的有用工具，以促进其DNA结合序列和相互作用蛋白质的共亲和纯化。此外，还报道了CrAsH-EDT2和N(1)-(4-叠氮基-2-硝基苯基)己烷-1,6-二胺合成方法的改进。两种光探针均能有效进入HeLa细胞（以及细胞核），并依赖重组DMRT1（双性和Mab3相关转录因子）中的四半胱氨酸基序来诱导荧光，这表明它们的交联能力可用于鉴定与感兴趣蛋白质相关的核酸和蛋白质。
