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

Sooim Shin, Moonsung Choi, Heather R Williamson, Victor L Davidson

Index: Biochim. Biophys. Acta 1837(10) , 1595-601, (2014)

Full Text: HTML

Abstract

The 6×-Histidine tag which is commonly used for purification of recombinant proteins was converted to a catalytic redox-active center by incorporation of Co(2+). 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 Co(2+)-loaded 6×His-tag could mediate long range electron transfer, it was shown that addition of H2O2 to the Co(2+)-loaded 6×His-tagged Cu(1+) amicyanin oxidizes the copper site which is 20Å 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. Copyright © 2014 Elsevier B.V. All rights reserved.