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Title: The creation of the artificial RING finger from the cross-brace zinc finger by {alpha}-helical region substitution

Abstract

The creation of the artificial RING finger as ubiquitin-ligating enzyme (E3) has been demonstrated. In this study, by the {alpha}-helical region substitution between the EL5 RING finger and the Williams-Beuren syndrome transcription factor (WSTF) PHD finger, the artificial E3 (WSTF PHD{sub R}ING finger) was newly created. The experiments of the chemical modification of residues Cys and the circular dichroism spectra revealed that the WSTF PHD{sub R}ING finger binds two zinc atoms and adopts the zinc-dependent ordered-structure. In the substrate-independent ubiquitination assay, the WSTF PHD{sub R}ING finger functions as E3 and was poly- or mono-ubiquitinated. The present strategy is very simple and convenient, and consequently it might be widely applicable to the creation of various artificial E3 RING fingers with the specific ubiquitin-conjugating enzyme (E2)-binding capability.

Authors:
 [1];  [2]
  1. Department of Pharmaceutical Health Care, Faculty of Pharmaceutical Sciences, Himeji Dokkyo University, Hyogo 670-8524 (Japan)
  2. 15-203 Hirano-machi, Himeji-shi, Hyogo 670-0933 (Japan)
Publication Date:
OSTI Identifier:
22202480
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 394; Journal Issue: 4; Other Information: Copyright (c) 2010 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; DICHROISM; ENZYMES; GENES; MALTOSE; MBP; MODIFICATIONS; RESIDUES; SPECTRA; SUBSTRATES; TRANSCRIPTION FACTORS

Citation Formats

Miyamoto, Kazuhide, and Togiya, Kayo. The creation of the artificial RING finger from the cross-brace zinc finger by {alpha}-helical region substitution. United States: N. p., 2010. Web. doi:10.1016/J.BBRC.2010.03.100.
Miyamoto, Kazuhide, & Togiya, Kayo. The creation of the artificial RING finger from the cross-brace zinc finger by {alpha}-helical region substitution. United States. https://doi.org/10.1016/J.BBRC.2010.03.100
Miyamoto, Kazuhide, and Togiya, Kayo. 2010. "The creation of the artificial RING finger from the cross-brace zinc finger by {alpha}-helical region substitution". United States. https://doi.org/10.1016/J.BBRC.2010.03.100.
@article{osti_22202480,
title = {The creation of the artificial RING finger from the cross-brace zinc finger by {alpha}-helical region substitution},
author = {Miyamoto, Kazuhide and Togiya, Kayo},
abstractNote = {The creation of the artificial RING finger as ubiquitin-ligating enzyme (E3) has been demonstrated. In this study, by the {alpha}-helical region substitution between the EL5 RING finger and the Williams-Beuren syndrome transcription factor (WSTF) PHD finger, the artificial E3 (WSTF PHD{sub R}ING finger) was newly created. The experiments of the chemical modification of residues Cys and the circular dichroism spectra revealed that the WSTF PHD{sub R}ING finger binds two zinc atoms and adopts the zinc-dependent ordered-structure. In the substrate-independent ubiquitination assay, the WSTF PHD{sub R}ING finger functions as E3 and was poly- or mono-ubiquitinated. The present strategy is very simple and convenient, and consequently it might be widely applicable to the creation of various artificial E3 RING fingers with the specific ubiquitin-conjugating enzyme (E2)-binding capability.},
doi = {10.1016/J.BBRC.2010.03.100},
url = {https://www.osti.gov/biblio/22202480}, journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 4,
volume = 394,
place = {United States},
year = {Fri Apr 16 00:00:00 EDT 2010},
month = {Fri Apr 16 00:00:00 EDT 2010}
}