Title

A Simple Method To Engineer A Protein-Derived Redox Cofactor For Catalysis

Keywords

Bioenergetics; Biotechnology; Enzyme; Histidine tag; Protein Engineering

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 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. © 2014 Elsevier B.V.

Publication Date

1-1-2014

Publication Title

Biochimica et Biophysica Acta - Bioenergetics

Volume

1837

Issue

10

Number of Pages

1595-1601

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1016/j.bbabio.2014.05.354

Socpus ID

84905780729 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/84905780729

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