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2hot

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|PDB= 2hot |SIZE=350|CAPTION= <scene name='initialview01'>2hot</scene>, resolution 2.19&Aring;
|PDB= 2hot |SIZE=350|CAPTION= <scene name='initialview01'>2hot</scene>, resolution 2.19&Aring;
|SITE=
|SITE=
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|LIGAND= <scene name='pdbligand=P2O:3-PROP-2-YN-1-YL-1,3-OXAZOLIDIN-2-ONE'>P2O</scene> and <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>
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|LIGAND= <scene name='pdbligand=DA:2&#39;-DEOXYADENOSINE-5&#39;-MONOPHOSPHATE'>DA</scene>, <scene name='pdbligand=DC:2&#39;-DEOXYCYTIDINE-5&#39;-MONOPHOSPHATE'>DC</scene>, <scene name='pdbligand=DG:2&#39;-DEOXYGUANOSINE-5&#39;-MONOPHOSPHATE'>DG</scene>, <scene name='pdbligand=DT:THYMIDINE-5&#39;-MONOPHOSPHATE'>DT</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=P2O:3-PROP-2-YN-1-YL-1,3-OXAZOLIDIN-2-ONE'>P2O</scene>
|ACTIVITY=
|ACTIVITY=
|GENE= En ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=7227 Drosophila melanogaster])
|GENE= En ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=7227 Drosophila melanogaster])
 +
|DOMAIN=
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|RELATEDENTRY=[[2hdd|2HDD]]
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|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2hot FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2hot OCA], [http://www.ebi.ac.uk/pdbsum/2hot PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=2hot RCSB]</span>
}}
}}
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[[Category: Shokat, K M.]]
[[Category: Shokat, K M.]]
[[Category: Simon, M D.]]
[[Category: Simon, M D.]]
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[[Category: GOL]]
 
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[[Category: P2O]]
 
[[Category: homeodomain]]
[[Category: homeodomain]]
[[Category: phage display]]
[[Category: phage display]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 17:19:36 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Mar 31 03:33:13 2008''

Revision as of 00:33, 31 March 2008


PDB ID 2hot

Drag the structure with the mouse to rotate
, resolution 2.19Å
Ligands: , , , , ,
Gene: En (Drosophila melanogaster)
Related: 2HDD


Resources: FirstGlance, OCA, PDBsum, RCSB
Coordinates: save as pdb, mmCIF, xml



Phage selected homeodomain bound to modified DNA


Overview

The homeodomain (HD)-DNA interface has been conserved over 500 million years of evolution. Despite this conservation, we have successfully re-engineered the engrailed HD to specifically recognize an unnatural nucleotide using a phage display selection. Here we report the synthesis of novel nucleosides and the selection of mutant HDs that bind these nucleotides using phage display. The high-resolution crystal structure of one mutant in complex with modified and unmodified DNA demonstrates that, even with the substantial perturbation to the interface, this selected mutant retains a canonical HD structure. Dissection of the contributions due to each of the selected mutations reveals that the majority of the modification-specific binding is accomplished by a single mutation (I47G) but that the remaining mutations retune the stability of the HD. These results afford a detailed look at a re-engineered protein-DNA interaction and provide insight into the opportunities for re-engineering highly conserved interfaces.

About this Structure

2HOT is a Single protein structure of sequence from Drosophila melanogaster. Full crystallographic information is available from OCA.

Reference

Structure and properties of a re-engineered homeodomain protein-DNA interface., Simon MD, Feldman ME, Rauh D, Maris AE, Wemmer DE, Shokat KM, ACS Chem Biol. 2006 Dec 15;1(12):755-60. PMID:17240973

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