2hos
Phage-Selected Homeodomain Bound to Unmodified DNAPhage-Selected Homeodomain Bound to Unmodified DNA
Structural highlights
FunctionHMEN_DROME This protein specifies the body segmentation pattern. It is required for the development of the central nervous system. Transcriptional regulator that represses activated promoters. Wg signaling operates by inactivating the SGG repression of EN autoactivation. Evolutionary Conservation Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf. Publication Abstract from PubMedThe 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. 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[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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