2hot: Difference between revisions
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[[Image:2hot.gif|left|200px]] | [[Image:2hot.gif|left|200px]] | ||
'''Phage selected homeodomain bound to modified DNA''' | {{Structure | ||
|PDB= 2hot |SIZE=350|CAPTION= <scene name='initialview01'>2hot</scene>, resolution 2.19Å | |||
|SITE= | |||
|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> | |||
|ACTIVITY= | |||
|GENE= En ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=7227 Drosophila melanogaster]) | |||
}} | |||
'''Phage selected homeodomain bound to modified DNA''' | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
2HOT is a [ | 2HOT is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Drosophila_melanogaster Drosophila melanogaster]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2HOT OCA]. | ||
==Reference== | ==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:[http:// | 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:[http://www.ncbi.nlm.nih.gov/pubmed/17240973 17240973] | ||
[[Category: Drosophila melanogaster]] | [[Category: Drosophila melanogaster]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
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[[Category: phage display]] | [[Category: phage display]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 17:19:36 2008'' | ||
Revision as of 15:19, 20 March 2008
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| 2hot, resolution 2.19Å | |||||||||||||
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| Ligands: | P2O and GOL | ||||||||||||
| Gene: | En (Drosophila melanogaster) | ||||||||||||
| 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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