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1se9

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[[Image:1se9.gif|left|200px]]<br /><applet load="1se9" size="350" color="white" frame="true" align="right" spinBox="true"
 
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caption="1se9" />
 
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'''Structure of At3g01050, a ubiquitin-fold protein from Arabidopsis thaliana'''<br />
 
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==Overview==
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==Structure of At3g01050, a ubiquitin-fold protein from Arabidopsis thaliana==
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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.
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<StructureSection load='1se9' size='340' side='right'caption='[[1se9]]' scene=''>
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== Structural highlights ==
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<table><tr><td colspan='2'>[[1se9]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Arabidopsis_thaliana Arabidopsis thaliana]. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1SE9 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1SE9 FirstGlance]. <br>
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</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Solution NMR</td></tr>
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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1se9 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1se9 OCA], [https://pdbe.org/1se9 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1se9 RCSB], [https://www.ebi.ac.uk/pdbsum/1se9 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1se9 ProSAT], [https://www.topsan.org/Proteins/CESG/1se9 TOPSAN]</span></td></tr>
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</table>
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== Function ==
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[https://www.uniprot.org/uniprot/MUB1_ARATH MUB1_ARATH] May serve as docking site to facilitate the association of other proteins to the plasma membrane.
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== Evolutionary Conservation ==
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[[Image:Consurf_key_small.gif|200px|right]]
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Check<jmol>
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<jmolCheckbox>
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<scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/se/1se9_consurf.spt"</scriptWhenChecked>
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<scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
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<text>to colour the structure by Evolutionary Conservation</text>
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</jmolCheckbox>
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</jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=1se9 ConSurf].
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<div style="clear:both"></div>
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<div style="background-color:#fffaf0;">
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== Publication Abstract from PubMed ==
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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.
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==About this Structure==
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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<ref>PMID:15782178</ref>
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1SE9 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Arabidopsis_thaliana Arabidopsis thaliana]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1SE9 OCA].
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==Reference==
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From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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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:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=15782178 15782178]
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</div>
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<div class="pdbe-citations 1se9" style="background-color:#fffaf0;"></div>
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== References ==
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<references/>
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__TOC__
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</StructureSection>
[[Category: Arabidopsis thaliana]]
[[Category: Arabidopsis thaliana]]
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[[Category: Single protein]]
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[[Category: Large Structures]]
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[[Category: CESG, Center.for.Eukaryotic.Structural.Genomics.]]
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[[Category: Lytle BL]]
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[[Category: Lytle, B.L.]]
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[[Category: Peterson FC]]
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[[Category: Peterson, F.C.]]
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[[Category: Volkman BF]]
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[[Category: Volkman, B.F.]]
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[[Category: cell-free]]
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[[Category: center for eukaryotic structural genomics]]
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[[Category: cesg]]
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[[Category: plant protein]]
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[[Category: protein structure initiative]]
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[[Category: psi]]
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[[Category: structural genomics]]
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[[Category: ubiquitin-like]]
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[[Category: wheat germ]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Feb 13 08:16:08 2008''
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Current revision

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

PDB ID 1se9

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