Jump to main content
Jump to site search

Issue 2, 2014
Previous Article Next Article

Constructing a man-made c-type cytochrome maquette in vivo: electron transfer, oxygen transport and conversion to a photoactive light harvesting maquette.

Author affiliations

Abstract

The successful use of man-made proteins to advance synthetic biology requires both the fabrication of functional artificial proteins in a living environment, and the ability of these proteins to interact productively with other proteins and substrates in that environment. Proteins made by the maquette method integrate sophisticated oxidoreductase function into evolutionarily naive, non-computationally designed protein constructs with sequences that are entirely unrelated to any natural protein. Nevertheless, we show here that we can efficiently interface with the natural cellular machinery that covalently incorporates heme into natural cytochromes c to produce in vivo an artificial c-type cytochrome maquette. Furthermore, this c-type cytochrome maquette is designed with a displaceable histidine heme ligand that opens to allow functional oxygen binding, the primary event in more sophisticated functions ranging from oxygen storage and transport to catalytic hydroxylation. To exploit the range of functions that comes from the freedom to bind a variety of redox cofactors within a single maquette framework, this c-type cytochrome maquette is designed with a second, non-heme C, tetrapyrrole binding site, enabling the construction of an elementary electron transport chain, and when the heme C iron is replaced with zinc to create a Zn porphyrin, a light-activatable artificial redox protein. The work we describe here represents a major advance in de novo protein design, offering a robust platform for new c-type heme based oxidoreductase designs and an equally important proof-of-principle that cofactor-equipped man-made proteins can be expressed in living cells, paving the way for constructing functionally useful man-made proteins in vivo.

Graphical abstract: Constructing a man-made c-type cytochrome maquette in vivo: electron transfer, oxygen transport and conversion to a photoactive light harvesting maquette.

Back to tab navigation

Supplementary files

Additions and corrections

Publication details

The article was received on 18 Jul 2013, accepted on 30 Oct 2013 and first published on 31 Oct 2013


Article type: Edge Article
DOI: 10.1039/C3SC52019F
Citation: Chem. Sci., 2014,5, 507-514
  •   Request permissions

    Constructing a man-made c-type cytochrome maquette in vivo: electron transfer, oxygen transport and conversion to a photoactive light harvesting maquette.

    J. L. R. Anderson, C. T. Armstrong, G. Kodali, B. R. Lichtenstein, D. W. Watkins, J. A. Mancini, A. L. Boyle, T. A. Farid, M. P. Crump, C. C. Moser and P. L. Dutton, Chem. Sci., 2014, 5, 507
    DOI: 10.1039/C3SC52019F

Search articles by author

Spotlight

Advertisements