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Identification and Characterization of a Redox Sensor Phosphodiesterase from Ferrovum sp. PN-J185 Containing Bacterial Hemerythrin and HD-GYP Domains
journal contribution
posted on 2020-02-20, 19:36 authored by Kenichi Kitanishi, Jotaro Igarashi, Ariki Matsuoka, Masaki UnnoThe second messenger
bis(3′,5′)-cyclic dimeric guanosine
monophosphate (c-di-GMP) regulates numerous important physiological
functions in bacteria. In this study, we identified and characterized
the first dimeric, full-length, non-heme iron-bound phosphodiesterase
(PDE) containing bacterial hemerythrin and HD-GYP domains (Bhr-HD-GYP).
We found that the amino acid sequence encoded by the FV185_09380 gene from Ferrovum sp. PN-J185 contains an N-terminal
bacterial hemerythrin domain and a C-terminal HD-GYP domain, which
is characteristic of proteins with PDE activity toward c-di-GMP. Inductively
coupled plasma optical emission spectroscopy analyses showed that
Bhr-HD-GYP contains 4 equiv of iron atoms per subunit, suggesting
both hemerythrin and HD-GYP domains have non-heme di-iron sites. A
redox-dependent spectral change expected for oxo-bridged non-heme
iron with carboxylate ligands was observed, and this redox interconversion
was reversible. However, unlike marine invertebrate hemerythrin, which
functions as an oxygen-binding protein, Bhr-HD-GYP did not form an
oxygen adduct because of rapid autoxidation. The reduced ferrous iron
complex of the protein catalyzed the hydrolysis of c-di-GMP to its
linearized product, 5′-phosphoguanylyl-(3′,5′)-guanosine
(pGpG), whereas the oxidized ferric iron complex had no significant
activity. These results suggest that Bhr-HD-GYP is a redox and oxygen
sensor enzyme that regulates c-di-GMP levels in response to changes
in cellular redox status or oxygen concentration. Our study may lead
to an improved understanding of the physiology of iron-oxidizing bacterium Ferrovum sp. PN-J185.
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non-heme di-iron sitesdimericiron-oxidizing bacterium Ferrovum spc-di-GMPemission spectroscopy analysesBhr-HD-GYPPN-J 185 Containingoxo-bridged non-heme ironnon-heme iron-bound phosphodiesteraseFVmarine invertebrate hemerythrinredoxHD-GYP domainsC-terminal HD-GYP domainfunctionRedox Sensor PhosphodiesterasePDEproteinoxygen sensor enzymeFerrovum sp
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