4dne: Difference between revisions
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[[ | ==Crystal structure of a triple-mutant of streptavidin in complex with desthiobiotin== | ||
<StructureSection load='4dne' size='340' side='right' caption='[[4dne]], [[Resolution|resolution]] 1.88Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[4dne]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Streptomyces_avidinii Streptomyces avidinii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4DNE OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4DNE FirstGlance]. <br> | |||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=DTB:6-(5-METHYL-2-OXO-IMIDAZOLIDIN-4-YL)-HEXANOIC+ACID'>DTB</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4dne FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4dne OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4dne RCSB], [http://www.ebi.ac.uk/pdbsum/4dne PDBsum]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Attempts to crystallize AtNTT1, a chloroplast ATP/ADP transporter from Arabidopsis thaliana, revealed an unexpected contaminant, Strep-Tactin, a variant of streptavidin that was used during purification of the protein. Although it was present in very small amounts, crystals of Strep-Tactin were reproducibly grown from the AtNTT1 solution. AtNTT1 was overexpressed in Escherichia coli and purified from detergent-solubilized membrane fractions using Strep-Tactin affinity chromatography based on an engineered streptavidin. The contamination of protein solutions purified on Strep-Tactin columns has never been described previously and seems to be specific to membrane proteins solubilized in detergents. Trace amounts of Strep-Tactin were observed to be eluted from a Strep-Tactin column using several routinely used detergents, illustrating their possible role in the contamination. This finding raises an alarm and suggests caution in membrane-protein purification using Strep-Tactin affinity columns, where detergents are essential components. The small crystals of contaminant protein led to the structure at 1.9 A resolution of Strep-Tactin in complex with desthiobiotin. | |||
Contamination from an affinity column: an encounter with a new villain in the world of membrane-protein crystallization.,Panwar P, Deniaud A, Pebay-Peyroula E Acta Crystallogr D Biol Crystallogr. 2012 Oct;68(Pt 10):1272-7. Epub 2012 Sep 13. PMID:22993081<ref>PMID:22993081</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
==See Also== | ==See Also== | ||
*[[Avidin|Avidin]] | *[[Avidin|Avidin]] | ||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Streptomyces avidinii]] | [[Category: Streptomyces avidinii]] | ||
[[Category: Deniaud, A | [[Category: Deniaud, A]] | ||
[[Category: Panwar, P | [[Category: Panwar, P]] | ||
[[Category: Pebay-Peyroula, E | [[Category: Pebay-Peyroula, E]] | ||
[[Category: Biotin]] | [[Category: Biotin]] | ||
[[Category: Biotin-binding protein]] | [[Category: Biotin-binding protein]] | ||
Revision as of 16:38, 9 December 2014
Crystal structure of a triple-mutant of streptavidin in complex with desthiobiotin
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