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

Shameem S. Syeda; Daren Rice; Derek J. Hook; Leslie L. Heckert; Gunda I. Georg

doi:10.1002/ardp.201500440

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

Shameem S. Syeda, Daren Rice, Derek J. Hook, Leslie L. Heckert, Gunda I. Georg

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

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 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.

Original language	English (US)
Pages (from-to)	233-241
Number of pages	9
Journal	Archiv der Pharmazie
Volume	349
Issue number	4
DOIs	https://doi.org/10.1002/ardp.201500440
State	Published - Apr 1 2016

Bibliographical note

Publisher Copyright:
© 2016 The Authors. Archiv der Pharmazie Published by Wiley-VCH Verlag GmbH & Co. KGaA.

Keywords

Biarsenical probe
Fluorescence
Photoaffinity probes
Tetracysteine-tagged recombinant DMRT1

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10.1002/ardp.201500440

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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 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.",

keywords = "Biarsenical probe, Fluorescence, Photoaffinity probes, Tetracysteine-tagged recombinant DMRT1",

author = "Syeda, {Shameem S.} and Daren Rice and Hook, {Derek J.} and Heckert, {Leslie L.} and Georg, {Gunda I.}",

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T1 - Synthesis of Arylazide- and Diazirine-Containing CrAsH-EDT2 Photoaffinity Probes

AU - Syeda, Shameem S.

AU - Rice, Daren

AU - Hook, Derek J.

AU - Heckert, Leslie L.

AU - Georg, Gunda I.

PY - 2016/4/1

Y1 - 2016/4/1

N2 - 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.

AB - 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.

KW - Biarsenical probe

KW - Fluorescence

KW - Photoaffinity probes

KW - Tetracysteine-tagged recombinant DMRT1

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