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2hot
From Proteopedia
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[[Image:2hot.gif|left|200px]] | [[Image:2hot.gif|left|200px]] | ||
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'''Phage selected homeodomain bound to modified DNA''' | '''Phage selected homeodomain bound to modified DNA''' | ||
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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: | + | [[Category: Phage display]] |
| - | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May 4 06:31:37 2008'' | |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | |
Revision as of 03:31, 4 May 2008
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 Page seeded by OCA on Sun May 4 06:31:37 2008
