Modular Artificial Cupredoxins

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• 作者：Samuel I. Mann ; Tillmann Heinisch ; Andrew C. Weitz ; Michael P. Hendrich ; Thomas R. Ward ; A. S. Borovik
• 刊名：Journal of the American Chemical Society
• 出版年：2016
• 出版时间：July 27, 2016
• 年：2016
• 卷：138
• 期：29
• 页码：9073-9076
• 全文大小：271K
• 年卷期：0
• ISSN：1520-5126

文摘

Cupredoxins are electron-transfer proteins that have active sites containing a mononuclear Cu center with an unusual trigonal monopyramidal structure (Type 1 Cu). A single Cu–S_cys bond is present within the trigonal plane that is responsible for its unique physical properties. We demonstrate that a cysteine-containing variant of streptavidin (Sav) can serve as a protein host to model the structure and properties of Type 1 Cu sites. A series of artificial Cu proteins are described that rely on Sav and a series of biotinylated synthetic Cu complexes. Optical and EPR measurements highlight the presence of a Cu–S_cys bond, and XRD analysis provides structural evidence. We further provide evidence that changes in the linker between the biotin and Cu complex within the synthetic constructs allows for small changes in the placement of Cu centers within Sav that have dramatic effects on the structural and physical properties of the resulting artificial metalloproteins. These findings highlight the utility of the biotin-Sav technology as an approach for simulating active sites of metalloproteins.