1SE9: Structure of At3g01050, a ubiquitin-fold protein from Arabidopsis thaliana

Citation:
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.
PDB ID: 1SE9Download
MMDB ID: 26628
PDB Deposition Date: 2004/2/16
Updated in MMDB: 2004/12
Experimental Method:
solution nmr
Source Organism:
Similar Structures:
Molecular Components in 1SE9
Label Count Molecule
Protein (1 molecule)
1
Ubiquitin Family(Gene symbol: MUB1)
Molecule annotation
* Click molecule labels to explore molecular sequence information.

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