1SE9

Structure of At3g01050, a ubiquitin-fold protein from Arabidopsis thaliana

1SE9 の概要

• エントリーDOI: 10.2210/pdb1se9/pdb
• NMR情報: BMRB: 6128
• 分子名称: ubiquitin family (1 entity in total)
• 機能のキーワード: ubiquitin-like, cell-free, wheat germ, structural genomics, protein structure initiative, cesg, psi, center for eukaryotic structural genomics, plant protein
• 由来する生物種: Arabidopsis thaliana (thale cress)
• 細胞内の位置: Cell membrane; Lipid-anchor: Q9MAB9
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 13970.13

構造登録者

Volkman, B.F.,Lytle, B.L.,Peterson, F.C.,Center for Eukaryotic Structural Genomics (CESG) (登録日: 2004-02-16, 公開日: 2004-02-24, 最終更新日: 2024-05-22)

主引用文献

Vinarov, D.A.,Lytle, B.L.,Peterson, F.C.,Tyler, E.M.,Volkman, B.F.,Markley, J.L.
Cell-free protein production and labeling protocol for NMR-based structural proteomics.
Nat.Methods, 1:149-153, 2004

PubMed Abstract: Structural proteomics requires robust, scalable methods. Here we describe a wheat germ cell-free platform for protein production that supports efficient NMR structural studies of eukaryotic proteins and offers advantages over cell-based methods. To illustrate this platform, we describe its application to a specific target (At3g01050.1) from Arabidopsis thaliana. After cloning the target gene into a specialized plasmid, we carry out a small-scale (50 mul) in vitro sequential transcription and translation trial to ascertain the level of protein production and solubility. Next, we prepare mRNA for use in a 4-ml semicontinuous cell-free translation reaction to incorporate (15)N-labeled amino acids into a protein sample that we purify and test for suitability for NMR structural analysis. We then repeat the cell-free approach with (13)C,(15)N-labeled amino acids to prepare a doubly labeled sample. The three-dimensional (3D) structure of At3g01050.1 shows that this protein is an unusual member of the beta-grasp protein family.

PubMed: 15782178
DOI: 10.1038/nmeth716
主引用文献が同じPDBエントリー

実験手法

SOLUTION NMR