A Simple Method to Engineer a Protein-Derived Redox Cofactor for Catalysis
National Institutes of Health, National Institute of General Medical Sciences
Department of Chemistry
The 6 ×-Histidine tag which is commonly used for purification of recombinant proteins was converted to a catalytic redox-active center by incorporation of Co2 +. Two examples of the biological activity of this engineered protein-derived cofactor are presented. After inactivation of the natural diheme cofactor of MauG, it was shown that the Co 2 +-loaded 6 × His-tag could substitute for the hemes in the H2O2-driven catalysis of tryptophan tryptophylquinone biosynthesis. To further demonstrate that the Co2 +-loaded 6 × His-tag could mediate long range electron transfer, it was shown that addition of H2O2 to the Co2 +-loaded 6 × His-tagged Cu1 + amicyanin oxidizes the copper site which is 20 Å away. These results provide proof of principle for this simple method by which to introduce a catalytic redox-active site into proteins for potential applications in research and biotechnology.
Shin, S.; Choi, M.; Williamson, Heather R.; and Davidson, V. L., "A Simple Method to Engineer a Protein-Derived Redox Cofactor for Catalysis" (2014). Faculty and Staff Publications. 157.