Iron-sufficient Azotobacter salinestris cells bound large amounts of 55Fe to cell-associated catechol melanin in an energy-independent manner. Iron was mobilized from the cell surface by citric acid and transported into the cell in a process that was inhibited by azide, carbonyl cyanide m-chlorophenyl-hydrazone (CCCP), KCl or RbCl, the latter two known to inhibit Na(+)-dependent activities in A. salinestris. Iron-limited cells produced a hydroxamate compound (HDX) which promoted 55Fe-uptake into iron-limited cells in a two step process. Initial uptake was inhibited by azide or CCCP, but not by KCl, while subsequent uptake was blocked by all inhibitors. Citric acid also mediated energy-dependent 55Fe-uptake in iron-limited cells, but initial iron-uptake was less sensitive to CCCP than HDX-mediated iron-uptake. The results show that melanin serves as an iron trap, probably to protect the cells from oxidative damage mediated by H2O2 and the Fenton reaction. A model for HDX siderophore-mediated iron-uptake is proposed which requires energy to concentrate iron in the periplasm and H+/Na(+)-dependent events to bring iron into the cell.
"The contribution of melanin to cell wall charge has been studied in Cryptococcus neoformans (Nosanchuk and Casadevall 1997). An investigation of iron binding by the melanin of a bacterium , Azotobacter salinestris (Page and Shivprasad 1995), may provide hints of a similar function for fungal melanins. These authors proposed that the bacterial melanin acts as an iron trap to protect cells from oxidative damage by hydrogen peroxide generated by iron-mediated Fenton reactions. "
[Show abstract][Hide abstract] ABSTRACT: The relationship of polyketide melanogenesis molecular biology to that of nonmelanin-producing pathways in a wide range of fungi and other organisms is discussed. Analytical methods and fundamental properties of melanins are discussed and fungal melanin properties are compared with those of animal and bacterial melanins. The enzymatic degradation of melanins by lignin peroxidases is described.Key words: fungal melanin, polyketide melanin, DHN melanin, melanin degradation, melanin properties, melanin analysis.
Canadian Journal of Microbiology 02/2011; 44(12):1115-1136. DOI:10.1139/w98-119 · 1.22 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Melanins represent virulence factors for several pathogenic fungi; the number of examples is growing. Thus, albino mutants of several genera (in one case, mutated precisely in the melanizing enzyme) exhibit decreased virulence in mice. We consider the phenomenon in relation to known chemical properties of melanin, beginning with biosynthesis from ortho-hydroquinone precursors which, when oxidized enzymatically to quinones, polymerize spontaneously to melanin. It follows that melanizing intermediates are cross-linking reagents; melanization stabilizes the external cell wall against hydrolysis and is thought to determine semipermeability in the osmotic ram (the appressorium) of certain plant pathogens. Polymeric melanins undergo reversible oxidation-reduction reactions between cell wall-penetrating quinone and hydroquinone oxidation states and thus represent polymeric redox buffers; using strong oxidants, it is possible to titrate the melanin on living cells and thereby demonstrate protection conferred by melanin in several species. The amount of buffering per cell approximately neutralizes the amount of oxidant generated by a single macrophage. Moreover, the intermediate oxidation state, the semiquinone, is a very stable free radical and is thought to trap unpaired electrons. We have suggested that the oxidation state of external melanin may be regulated by external Fe(II). An independent hypothesis holds that in Cryptococcus neoformans, an important function of the melanizing enzyme (apart from melanization) is the oxidation of Fe(II) to Fe(III), thereby forestalling generation of the harmful hydroxyl radical from H(2)O(2). Thus, problems in fungal pathogenesis have led to evolving hypotheses regarding melanin functioning.
[Show abstract][Hide abstract] ABSTRACT: Melanins are enigmatic pigments that are produced by a wide variety of microorganisms including several species of pathogenic bacteria, fungi and helminths. The study of melanin is difficult because these pigments defy complete biochemical and structural analysis. Nevertheless, the availability of new reagents in the form of monoclonal antibodies and melanin-binding peptides, combined with the application of various physical techniques, has provided insights into the process of melanization. Melanization is important in microbial pathogenesis because it has been associated with virulence in many microorganisms. Melanin appears to contribute to virulence by reducing the susceptibility of melanized microbes to host defence mechanisms. However, the interaction of melanized microbes and the host is complex and includes immune responses to melanin-related antigens. Production of melanin has also been linked to protection against environmental insults. Interference with melanization is a potential strategy for antimicrobial drug and pesticide development. The process of melanization poses fascinating problems in cell biology and provides a type of pathogenic strategy that is common to highly diverse pathogens.
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