4O60

Structure of ankyrin repeat protein

4O60 の概要

• エントリーDOI: 10.2210/pdb4o60/pdb
• 関連するPDBエントリー: 4QFV
• 分子名称: ANK-N5C-317 (2 entities in total)
• 機能のキーワード: ankyrin, designed ankyrin repeats, de novo protein
• 由来する生物種: synthetic construct
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 50447.65

構造登録者

Ethayathulla, A.S.,Guan, L. (登録日: 2013-12-20, 公開日: 2015-03-25, 最終更新日: 2023-09-20)

主引用文献

Tikhonova, E.B.,Ethayathulla, A.S.,Su, Y.,Hariharan, P.,Xie, S.,Guan, L.
A transcription blocker isolated from a designed repeat protein combinatorial library by in vivo functional screen.
Sci Rep, 5:8070-8070, 2015

PubMed Abstract: A highly diverse DNA library coding for ankyrin seven-repeat proteins (ANK-N5C) was designed and constructed by a PCR-based combinatorial assembly strategy. A bacterial melibiose fermentation assay was adapted for in vivo functional screen. We isolated a transcription blocker that completely inhibits the melibiose-dependent expression of α-galactosidase (MelA) and melibiose permease (MelB) of Escherichia coli by specifically preventing activation of the melAB operon. High-resolution crystal structural determination reveals that the designed ANK-N5C protein has a typical ankyrin fold, and the specific transcription blocker, ANK-N5C-281, forms a domain-swapped dimer. Functional tests suggest that the activity of MelR, a DNA-binding transcription activator and a member of AraC family of transcription factors, is inhibited by ANK-N5C-281 protein. All ANK-N5C proteins are expected to have a concave binding area with negative surface potential, suggesting that the designed ANK-N5C library proteins may facilitate the discovery of binders recognizing structural motifs with positive surface potential, like in DNA-binding proteins. Overall, our results show that the established library is a useful tool for the discovery of novel bioactive reagents.

PubMed: 25627011
DOI: 10.1038/srep08070

実験手法

X-RAY DIFFRACTION (2.52 Å)