Consolidating critical binding determinants by noncyclic rearrangement of protein secondary structure
| Autor: | Ramon K. Tabtiang, Robert A. Grant, Brent O. Cezairliyan, Jesse C. Cochrane, Robert T. Sauer |
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| Rok vydání: | 2005 |
| Předmět: |
Models
Molecular Protein Folding Dimer Protein design Repressor Plasma protein binding Biology Crystallography X-Ray Protein Structure Secondary Viral Proteins chemistry.chemical_compound Viral Regulatory and Accessory Proteins Protein secondary structure Bacteriophage P22 Multidisciplinary Biological Sciences Recombinant Proteins Repressor Proteins Kinetics Crystallography chemistry DNA Viral Protein folding Linker DNA Protein Binding |
| Zdroj: | Proceedings of the National Academy of Sciences. 102:2305-2309 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.0409562102 |
| Popis: | We designed a single-chain variant of the Arc repressor homodimer in which the β strands that contact operator DNA are connected by a hairpin turn and the α helices that form the tetrahelical scaffold of the dimer are attached by a short linker. The designed protein represents a noncyclic permutation of secondary structural elements in another single-chain Arc molecule (Arc-L1-Arc), in which the two subunits are fused by a single linker. The permuted protein binds operator DNA with nanomolar affinity, refolds on the sub-millisecond time scale, and is as stable as Arc-L1-Arc. The crystal structure of the permuted protein reveals an essentially wild-type fold, demonstrating that crucial folding information is not encoded in the wild-type order of secondary structure. Noncyclic rearrangement of secondary structure may allow grouping of critical active-site residues in other proteins and could be a useful tool for protein design and minimization. |
| Databáze: | OpenAIRE |
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