Cu(II)-binding properties of a cytochrome c with a synthetic metal-binding site: His-X3-His in an alpha-helix
| Autor: | Frances H. Arnold, Barry L. Haymore, Robert J. Todd, Danilo R. Casimiro, Mariana E. Van Dam |
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| Rok vydání: | 1991 |
| Předmět: |
Hemeprotein
Saccharomyces cerevisiae Proteins Stereochemistry Protein Conformation Binding energy Molecular Sequence Data Metal Binding Site Cytochrome c Group Biochemistry Metal Structural Biology Enzyme Stability Histidine Amino Acid Sequence Binding site Molecular Biology biology Base Sequence Chemistry Cytochrome c Cytochromes c Genetic Variation Stability constants of complexes visual_art Helix visual_art.visual_art_medium biology.protein Copper Protein Binding |
| Zdroj: | Proteins. 10(2) |
| ISSN: | 0887-3585 |
| Popis: | A metal-binding site consisting of two histidines positioned His-X3-His in an alpha-helix has been engineered into the surface of Saccharomyces cerevisiae iso-1-cytochrome c. The synthetic metal-binding cytochrome c retains its biological activity in vivo. Its ability to bind chelated Cu(II) has been characterized by partitioning in aqueous two-phase polymer systems containing a polymer-metal complex, Cu(II)IDA-PEG, and by metal-affinity chromatography. The stability constant for the complex formed between Cu(II)IDA-PEG and the cytochrome c His-X3-His site is 5.3 x 10(4) M-1, which corresponds to a chelate effect that contributes 1.5 kcal mol-1 to the binding energy. Incorporation of the His-X3-His site yields a synthetic metal-binding protein whose metal affinity is sensitive to environmental conditions that alter helix structure or flexibility. |
| Databáze: | OpenAIRE |
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