1se9: Difference between revisions
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|GENE= At3g01050 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=3702 Arabidopsis thaliana]) | |GENE= At3g01050 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=3702 Arabidopsis thaliana]) | ||
|DOMAIN=<span class='plainlinks'>[http://www.ncbi.nlm.nih.gov/Structure/cdd/cddsrv.cgi?uid=cd01814 NTGP5]</span> | |||
|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.fli-leibniz.de/cgi-bin/ImgLib.pl?CODE=1kfv JenaLib], [http://www.rcsb.org/pdb/explore.do?structureId=1se9 RCSB]</span> | |||
}} | }} | ||
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[[Category: wheat germ]] | [[Category: wheat germ]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Mar 26 06:04:39 2008'' | ||
Revision as of 04:04, 26 March 2008
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| Gene: | At3g01050 (Arabidopsis thaliana) | ||||||||||||
| Domains: | NTGP5 | ||||||||||||
| Resources: | FirstGlance, OCA, PDBsum, JenaLib, RCSB | ||||||||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||||||||
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
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