A simple method to engineer a protein-derived redox cofactor for catalysis

作者：Shin Sooim; Choi Moonsung; Williamson Heather R; Davidson Victor L^*

来源：Biochimica et Biophysica Acta-Bioenergetics, 2014, 1837(10): 1595-1601.

DOI：10.1016/j.bbabio.2014.05.354

• Protein Engineering
• Enzyme
• Biotechnology
• Bioenergetics
• Histidine tag

The 6x-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 Co2+-loaded 6x His-tag could substitute for the hemes in the H2O2-driven catalysis of tryptophan tryptophylquinone biosynthesis. To further demonstrate that the Co2+-loaded 6x His-tag could mediate long range electron transfer, it was shown that addition of H2O2 to the Co2+-loaded 6 x His-tagged Cu1+ amicyanin oxidizes the copper site which is 20 A 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.

• 出版日期2014-10