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

    Authors

    S. Shin; M. Choi; H. R. Williamson;V. L. Davidson

    Comments

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    Abbreviated Journal Title

    Biochim. Biophys. Acta-Bioenerg.

    Keywords

    Protein Engineering; Enzyme; Biotechnology; Bioenergetics; Histidine tag; TRYPTOPHAN TRYPTOPHYLQUINONE BIOSYNTHESIS; PARACOCCUS-DENITRIFICANS; METHYLAMINE DEHYDROGENASE; ELECTRON-TRANSFER; DESIGN; MAUG; COMPLEX; INTERMEDIATE; PURIFICATION; MUTAGENESIS; Biochemistry & Molecular Biology; Biophysics

    Abstract

    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. (C) 2014 Elsevier B.V. All rights reserved.

    Journal Title

    Biochimica Et Biophysica Acta-Bioenergetics

    Volume

    1837

    Issue/Number

    10

    Publication Date

    1-1-2014

    Document Type

    Article

    Language

    English

    First Page

    1595

    Last Page

    1601

    WOS Identifier

    WOS:000342273200001

    ISSN

    0005-2728

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