1SE9
Structure of At3g01050, a ubiquitin-fold protein from Arabidopsis thaliana
Summary for 1SE9
| Entry DOI | 10.2210/pdb1se9/pdb |
| NMR Information | BMRB: 6128 |
| Descriptor | ubiquitin family (1 entity in total) |
| Functional Keywords | ubiquitin-like, cell-free, wheat germ, structural genomics, protein structure initiative, cesg, psi, center for eukaryotic structural genomics, plant protein |
| Biological source | Arabidopsis thaliana (thale cress) |
| Cellular location | Cell membrane; Lipid-anchor: Q9MAB9 |
| Total number of polymer chains | 1 |
| Total formula weight | 13970.13 |
| Authors | Volkman, B.F.,Lytle, B.L.,Peterson, F.C.,Center for Eukaryotic Structural Genomics (CESG) (deposition date: 2004-02-16, release date: 2004-02-24, Last modification date: 2024-05-22) |
| Primary citation | 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 Cited by 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: 15782178DOI: 10.1038/nmeth716 PDB entries with the same primary citation |
| Experimental method | SOLUTION NMR |
Structure validation
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