2hot
From Proteopedia
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|PDB= 2hot |SIZE=350|CAPTION= <scene name='initialview01'>2hot</scene>, resolution 2.19Å | |PDB= 2hot |SIZE=350|CAPTION= <scene name='initialview01'>2hot</scene>, resolution 2.19Å | ||
|SITE= | |SITE= | ||
| - | |LIGAND= <scene name='pdbligand= | + | |LIGAND= <scene name='pdbligand=DA:2'-DEOXYADENOSINE-5'-MONOPHOSPHATE'>DA</scene>, <scene name='pdbligand=DC:2'-DEOXYCYTIDINE-5'-MONOPHOSPHATE'>DC</scene>, <scene name='pdbligand=DG:2'-DEOXYGUANOSINE-5'-MONOPHOSPHATE'>DG</scene>, <scene name='pdbligand=DT:THYMIDINE-5'-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= | ||
| + | |RELATEDENTRY=[[2hdd|2HDD]] | ||
| + | |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.]] | ||
| - | [[Category: GOL]] | ||
| - | [[Category: P2O]] | ||
[[Category: homeodomain]] | [[Category: homeodomain]] | ||
[[Category: phage display]] | [[Category: phage display]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | ''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
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| , 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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