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PDBsum entry 2hos
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Transcription/DNA
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PDB id
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2hos
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References listed in PDB file
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Key reference
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Title
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Structure and properties of a re-Engineered homeodomain protein-Dna interface.
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Authors
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M.D.Simon,
M.E.Feldman,
D.Rauh,
A.E.Maris,
D.E.Wemmer,
K.M.Shokat.
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Ref.
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Acs Chem Biol, 2006,
1,
755-760.
[DOI no: ]
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PubMed id
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Abstract
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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.
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