1se9: Difference between revisions

Revision as of 13:21, 30 April 2008

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

OverviewOverview

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 StructureAbout this Structure

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

ReferenceReference

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

Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)

OCA

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 [[Image:1se9.gif|left|200px]]
- {{Structure
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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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 '''Structure of At3g01050, a ubiquitin-fold protein from Arabidopsis thaliana'''
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 [[Category: Peterson, F C.]]
 [[Category: Volkman, B F.]]
-[[Category: cell-free]]
+[[Category: Cell-free]]
-[[Category: center for eukaryotic structural genomic]]
+[[Category: Center for eukaryotic structural genomic]]
-[[Category: cesg]]
+[[Category: Cesg]]
-[[Category: plant protein]]
+[[Category: Plant protein]]
-[[Category: protein structure initiative]]
+[[Category: Protein structure initiative]]
-[[Category: psi]]
+[[Category: Psi]]
-[[Category: structural genomic]]
+[[Category: Structural genomic]]
-[[Category: ubiquitin-like]]
+[[Category: Ubiquitin-like]]
-[[Category: wheat germ]]
+[[Category: Wheat germ]]
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Apr 30 13:21:04 2008''
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar 30 23:40:52 2008''