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

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

Biochimica et biophysica acta (2014-05-27)

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

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.

MATERIALS

Product Number

Brand

Product Description

Supelco

Tryptophan, Pharmaceutical Secondary Standard; Certified Reference Material

Sigma-Aldrich

Cobalt, foil, thickness 0.1 mm, 99.95% trace metals basis

Sigma-Aldrich

Cobalt, rod, diam. 5.0 mm, 99.95% trace metals basis

Sigma-Aldrich

Cobalt, foil, thickness 1.0 mm, 99.95% trace metals basis

Sigma-Aldrich

Cobalt, foil, thickness 0.1 mm, ≥99.99%

Sigma-Aldrich

Cobalt, wire, diam. 1.0 mm, 99.995% trace metals basis

Sigma-Aldrich

Cobalt, granular, 99.99% trace metals basis

Cobalt, foil, 25x25mm, thickness 1.0mm, as rolled, 99.99+%

Cobalt, foil, not light tested, 50x50mm, thickness 0.025mm, 99.9%

Sigma-Aldrich

Cobalt, pieces, 99.5% trace metals basis

Sigma-Aldrich

Cobalt, Carbon coated magnetic, nanopowder, <50 nm particle size (TEM), ≥99%

Cobalt, wire reel, 0.5m, diameter 0.5mm, hard, 99.99+%

Sigma-Aldrich

Cobalt, powder, 2 μm particle size, 99.8% trace metals basis

Cobalt, wire reel, 2m, diameter 0.05mm, hard, 99.9%

Cobalt, foil, not light tested, 25x25mm, thickness 0.008mm, 99.9%

Cobalt, foil, 8mm disks, thickness 0.025mm, 99.9%

Cobalt, foil, 8mm disks, thickness 0.007mm, 99.9%

Cobalt, foil, 8mm disks, thickness 0.05mm, as rolled, 99.99+%

Cobalt, foil, 6mm disks, thickness 0.006mm, 99.9%

Cobalt, foil, not light tested, 25x25mm, thickness 0.005mm, 99.9%

Cobalt, foil, not light tested, 50x50mm, thickness 0.0125mm, 99.9%

Cobalt, foil, 4mm disks, thickness 0.125mm, annealed, 99.9%

Cobalt, foil, 4mm disks, thickness 0.025mm, 99.9%

Cobalt, microfoil, disks, 10mm, thinness 0.25μm, specific density 222μg/cm², permanent mylar 3.5μm support, 99.9%

Cobalt, foil, 6mm disks, thickness 0.007mm, 99.9%

Cobalt, foil, 8mm disks, thickness 0.25mm, as rolled, 99.9%

Cobalt, foil, 8mm disks, thickness 0.05mm, as rolled, 99.9%

Cobalt, foil, light tested, 25x25mm, thickness 0.015mm, 99.9%

Cobalt, wire reel, 0.025m, diameter 2.0mm, hard, 99.99+%

Cobalt, foil, light tested, 100x100mm, thickness 0.05mm, as rolled, 99.99+%