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_RING finger) was newly created. The experiments of the chemical modification of residues Cys and the circular dichroism spectra revealed that the WSTF PHD_RING finger binds two zinc atoms and adopts the zinc-dependent ordered-structure. In the substrate-independent ubiquitination assay, the WSTF PHD_RING 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: Kazuhide Miyamoto, Kayo Togiya
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 394 Issue 4 Pg. 972-5 (Apr 16 2010) ISSN: 1090-2104 [Electronic] United States
• PMID: 20307496 (Publication Type: Journal Article)
• Copyright: 2010 Elsevier Inc. All rights reserved.

Chemical References

• BAZ1B protein, human
• Transcription Factors
• Ubiquitin-Protein Ligases
• Zinc
• Cysteine

Topics

• Amino Acid Sequence
• Cysteine (chemistry)
• Humans
• Molecular Sequence Data
• Protein Folding
• Protein Structure, Secondary
• RING Finger Domains
• Transcription Factors (chemistry)
• Ubiquitin-Protein Ligases (chemical synthesis, chemistry)
• Ubiquitination
• Zinc (chemistry)
• Zinc Fingers