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1 A single mutation within a flavoprotein is capable of switching the catalytic activity of a dehalogenase into a nitroreductase. This change in function correlates with a destabilization of the one-electron-reduced flavin semiquinone that is differentially expressed in the nitro-FMN reductase superfamily during redox cycling. The diversity of function within such a superfamily therefore has the potential to arise from rapid evolution, and its members should provide a convenient basis for developing new catalysts with an altered specificity of choice. 2 The coordination bond between gold and sulfur (Au¨CS) has been widely studied and utilized in many fields. However, detailed investigations on the basic nature of this bond are still lacking. A gold-specific binding protein, GolB, was recently identified, providing a unique opportunity for the study of the Au¨CS bond at the molecular level. We probed the mechanical strength of the gold¨Csulfur bond in GolB using single-molecule force spectroscopy. We measured the rupture force of the Au¨CS bond to be 165 pN, much lower than Au¨CS bonds measured on different gold surfaces (∼1000 pN). We further solved the structures of apo-GolB and Au(I)¨CGolB complex using X-ray crystallography. These structures showed that the average Au¨CS bond length in GolB is much longer than the reported average value of Au¨CS bonds. Our results highlight the dramatic influence of the unique biological environment on the stability and strength of metal coordination bonds in proteins. |
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