3zmf: Difference between revisions
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[[ | ==Salmonella enterica SadA 303-358 fused to GCN4 adaptors (SadAK2)== | ||
<StructureSection load='3zmf' size='340' side='right' caption='[[3zmf]], [[Resolution|resolution]] 1.85Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[3zmf]] is a 3 chain structure with sequence from [http://en.wikipedia.org/wiki/Salty Salty]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3ZMF OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3ZMF FirstGlance]. <br> | |||
</td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[2yny|2yny]], [[2ynz|2ynz]], [[2yo0|2yo0]], [[2yo1|2yo1]], [[2yo2|2yo2]], [[2yo3|2yo3]]</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=3zmf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3zmf OCA], [http://www.rcsb.org/pdb/explore.do?structureId=3zmf RCSB], [http://www.ebi.ac.uk/pdbsum/3zmf PDBsum]</span></td></tr> | |||
</table> | |||
== Function == | |||
[[http://www.uniprot.org/uniprot/GCN4_YEAST GCN4_YEAST]] Is a transcription factor that is responsible for the activation of more than 30 genes required for amino acid or for purine biosynthesis in response to amino acid or purine starvation. Binds and recognize the DNA sequence: 5'-TGA[CG]TCA-3'. | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
We repeatedly experienced difficulties in obtaining pure protein of a defined oligomeric state when expressing domains that consist partially or entirely of coiled coils. We therefore modified an established expression vector, pASK-IBA, to generate N- and C-terminal fusions of the cloned domain in heptad register with the GCN4 leucine zipper. GCN4 is a well-characterized coiled coil, for which stable dimeric, trimeric and tetrameric forms exist. To test this expression system, we produced a series of constructs derived from the trimeric autotransporter adhesin STM3691 of Salmonella (SadA), which has a highly repetitive structure punctuated by coiled-coil regions. The constructs begin and end with predicted coiled-coil segments of SadA, each fused in the correct heptad register to the trimeric form of GCN4, GCN4pII. All constructs were expressed at high levels, trimerized either natively or after refolding from inclusion bodies, and yielded crystals that diffracted to high resolution. Thus, fusion to GCN4pII allows for the efficient expression and crystallization of proteins containing trimeric coiled coils. The structure of short constructs can be solved conveniently by molecular replacement using the known GCN4 structure as a search model. The system can be adapted for constructs with dimeric or tetrameric coiled coils, using the corresponding GCN4 variants. | |||
A new expression system for protein crystallization using trimeric coiled-coil adaptors.,Hernandez Alvarez B, Hartmann MD, Albrecht R, Lupas AN, Zeth K, Linke D Protein Eng Des Sel. 2008 Jan;21(1):11-8. Epub 2007 Dec 18. PMID:18093992<ref>PMID:18093992</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
==See Also== | |||
*[[Gcn4|Gcn4]] | |||
== | == References == | ||
[[ | <references/> | ||
__TOC__ | |||
== | </StructureSection> | ||
< | [[Category: Salty]] | ||
[[Category: Albrecht, R | [[Category: Albrecht, R]] | ||
[[Category: Alvarez, B Hernandez | [[Category: Alvarez, B Hernandez]] | ||
[[Category: Hartmann, M D | [[Category: Hartmann, M D]] | ||
[[Category: Lupas, A N | [[Category: Lupas, A N]] | ||
[[Category: Dall domain]] | [[Category: Dall domain]] | ||
[[Category: Dall2]] | [[Category: Dall2]] | ||
[[Category: Membrane protein]] | [[Category: Membrane protein]] | ||
[[Category: Taa]] | [[Category: Taa]] | ||