1se9

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[[Image:1se9.gif|left|200px]]
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{{Structure
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|PDB= 1se9 |SIZE=350|CAPTION= <scene name='initialview01'>1se9</scene>
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The line below this paragraph, containing "STRUCTURE_1se9", creates the "Structure Box" on the page.
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|GENE= At3g01050 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=3702 Arabidopsis thaliana])
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|DOMAIN=<span class='plainlinks'>[http://www.ncbi.nlm.nih.gov/Structure/cdd/cddsrv.cgi?uid=cd01814 NTGP5]</span>
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{{STRUCTURE_1se9| PDB=1se9 | SCENE= }}
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|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=1se9 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1se9 OCA], [http://www.ebi.ac.uk/pdbsum/1se9 PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=1se9 RCSB]</span>
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'''Structure of At3g01050, a ubiquitin-fold protein from Arabidopsis thaliana'''
'''Structure of At3g01050, a ubiquitin-fold protein from Arabidopsis thaliana'''
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[[Category: Peterson, F C.]]
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[[Category: center for eukaryotic structural genomic]]
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[[Category: plant protein]]
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[[Category: Plant protein]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Apr 30 13:21:04 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar 30 23:40:52 2008''
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Revision as of 10:21, 30 April 2008

Image:1se9.gif

Template:STRUCTURE 1se9

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


Overview

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.

About this Structure

1SE9 is a Single protein structure of sequence from Arabidopsis thaliana. Full crystallographic information is available from OCA.

Reference

Cell-free protein production and labeling protocol for NMR-based structural proteomics., Vinarov DA, Lytle BL, Peterson FC, Tyler EM, Volkman BF, Markley JL, Nat Methods. 2004 Nov;1(2):149-53. Epub 2004 Oct 21. PMID:15782178 Page seeded by OCA on Wed Apr 30 13:21:04 2008

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