5LOZ

STRUCTURE OF YEAST ENT1 ENTH DOMAIN

5LOZ の概要

• エントリーDOI: 10.2210/pdb5loz/pdb
• 関連するPDBエントリー: 1EDU 1EYH 1HOA
• 分子名称: Epsin-1 (2 entities in total)
• 機能のキーワード: alpha-alpha superhelix, ubiquitin receptor, endocytosis adaptor, ubiquitin, clathrin, lipid, eps15, ubiquitin-binding domain
• 由来する生物種: Saccharomyces cerevisiae (Baker's yeast)
• 細胞内の位置: Cytoplasm: Q12518
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 15981.05

構造登録者

Tanner, N.,Prag, G. (登録日: 2016-08-11, 公開日: 2016-10-05, 最終更新日: 2024-05-08)

主引用文献

Levin-Kravets, O.,Tanner, N.,Shohat, N.,Attali, I.,Keren-Kaplan, T.,Shusterman, A.,Artzi, S.,Varvak, A.,Reshef, Y.,Shi, X.,Zucker, O.,Baram, T.,Katina, C.,Pilzer, I.,Ben-Aroya, S.,Prag, G.
A bacterial genetic selection system for ubiquitylation cascade discovery.
Nat.Methods, 13:945-952, 2016

PubMed Abstract: About one-third of the eukaryotic proteome undergoes ubiquitylation, but the enzymatic cascades leading to substrate modification are largely unknown. We present a genetic selection tool that utilizes Escherichia coli, which lack deubiquitylases, to identify interactions along ubiquitylation cascades. Coexpression of split antibiotic resistance protein tethered to ubiquitin and ubiquitylation target together with a functional ubiquitylation apparatus results in a covalent assembly of the resistance protein, giving rise to bacterial growth on selective media. We applied the selection system to uncover an E3 ligase from the pathogenic bacteria EHEC and to identify the epsin ENTH domain as an ultraweak ubiquitin-binding domain. The latter was complemented with a structure-function analysis of the ENTH-ubiquitin interface. We also constructed and screened a yeast fusion library, discovering Sem1 as a novel ubiquitylation substrate of Rsp5 E3 ligase. Collectively, our selection system provides a robust high-throughput approach for genetic studies of ubiquitylation cascades and for small-molecule modulator screening.

PubMed: 27694912
DOI: 10.1038/nmeth.4003

実験手法

X-RAY DIFFRACTION (1.95 Å)