2hos
From Proteopedia
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==Overview== | ==Overview== | ||
| - | The homeodomain (HD)-DNA interface has been conserved over 500 million | + | 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== | ==About this Structure== | ||
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[[Category: Drosophila melanogaster]] | [[Category: Drosophila melanogaster]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Feldman, M | + | [[Category: Feldman, M E.]] |
| - | [[Category: Shokat, K | + | [[Category: Shokat, K M.]] |
| - | [[Category: Simon, M | + | [[Category: Simon, M D.]] |
[[Category: 3MO]] | [[Category: 3MO]] | ||
[[Category: GOL]] | [[Category: GOL]] | ||
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[[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 Thu Feb 21 17:44:04 2008'' |
Revision as of 15:44, 21 February 2008
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Phage-Selected Homeodomain Bound to Unmodified 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
2HOS is a Single protein structure of sequence from Drosophila melanogaster with and as ligands. 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
Page seeded by OCA on Thu Feb 21 17:44:04 2008
