Design, Synthesis, and Characterization of a Photoactivatable Flavocytochrome Molecular Maquette

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1998 Sep 2

PMID 9724726

Citations 20

Authors

R E Sharp

C C Moser

F Rabanal

P L Dutton

Affiliations

Soon will be listed here.

Abstract

We report the construction of a synthetic flavo-heme protein that incorporates two major physiological activities of flavoproteins: light activation of flavin analogous to DNA photolyase and rapid intramolecular electron transfer between the flavin and heme cofactors as in several oxidoreductases. The functional tetra-alpha-helix protein comprises two 62-aa helix-loop-helix subunits. Each subunit contains a single cysteine to which flavin (7-acetyl-10-methylisoalloxazine) is covalently attached and two histidines appropriately positioned for bis-his coordination of heme cofactors. Both flavins and hemes are situated within the hydrophobic core of the protein. Intramolecular electron transfer from flavosemiquinone generated by photoreduction from a sacrificial electron donor in solution was examined between protoporphyrin IX and 1-methyl-2-oxomesoheme XIII. Laser pulse-activated electron transfer from flavin to meso heme occurs on a 100-ns time scale, with a favorable free energy of approximately -100 meV. Electron transfer from flavin to the lower potential protoporphyrin IX, with an unfavorable free energy, can be induced after a lag phase under continuous light illumination. Thus, the supporting peptide matrix provides an excellent framework for the positioning of closely juxtaposed redox groups capable of facilitating intramolecular electron transfer and begins to clarify in a simplified and malleable system the natural engineering of flavoproteins.

Citing Articles

Rational Construction of Compact de Novo-Designed Biliverdin-Binding Proteins.

Sheehan M, Magaraci M, Kuznetsov I, Mancini J, Kodali G, Moser C Biochemistry. 2018; 57(49):6752-6756.

PMID: 30468389 PMC: 6293442. DOI: 10.1021/acs.biochem.8b01076.

Design and engineering of water-soluble light-harvesting protein maquettes.

Kodali G, Mancini J, Solomon L, Episova T, Roach N, Hobbs C Chem Sci. 2017; 8(1):316-324.

PMID: 28261441 PMC: 5330312. DOI: 10.1039/C6SC02417C.

De Novo Construction of Redox Active Proteins.

Moser C, Sheehan M, Ennist N, Kodali G, Bialas C, Englander M Methods Enzymol. 2016; 580:365-88.

PMID: 27586341 PMC: 5123760. DOI: 10.1016/bs.mie.2016.05.048.

First principles design of a core bioenergetic transmembrane electron-transfer protein.

Goparaju G, Fry B, Chobot S, Wiedman G, Moser C, Dutton P Biochim Biophys Acta. 2015; 1857(5):503-512.

PMID: 26672896 PMC: 4846532. DOI: 10.1016/j.bbabio.2015.12.002.

Design and engineering of a man-made diffusive electron-transport protein.

Fry B, Solomon L, Dutton P, Moser C Biochim Biophys Acta. 2015; 1857(5):513-521.

PMID: 26423266 PMC: 4910091. DOI: 10.1016/j.bbabio.2015.09.008.

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