Stable, monomeric variants of lambda Cro obtained by insertion of a designed beta-hairpin sequence |
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Authors: | M C Mossing R T Sauer |
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Affiliation: | Department of Biology, Massachusetts Institute of Technology, Cambridge 02139. |
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Abstract: | lambda Cro is a dimeric DNA binding protein. Random mutagenesis and a selection for Cro activity have been used to identify the contacts between Cro subunits that are crucial for maintenance of a stably folded structure. To obtain equivalent contacts in a monomeric system, a Cro variant was designed and constructed in which the antiparallel beta-ribbon that forms the dimer interface was replaced by a beta-hairpin. The engineered monomer has a folded structure similar to wild type, is significantly more stable than wild type, and exhibits novel half-operator binding activity. |
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