"Fifty Shades" of Black and Red or How Carboxyl Groups Fine Tune Eumelanin and Pheomelanin Properties

Abstract

Recent advances in the chemistry of melanins have begun to disclose a number of important structure-property-function relationships of crucial relevance to the biological role of human pigments, including skin (photo) protection and UV-susceptibility. Even slight variations in the monomer composition of black eumelanins and red pheomelanins have been shown to determine significant differences in light absorption, antioxidant, paramagnetic and redox behavior, particle morphology, surface properties, metal chelation and resistance to photo-oxidative wear-and-tear. These variations are primarily governed by the extent of decarboxylation at critical branching points of the eumelanin and pheomelanin pathways, namely the rearrangement of dopachrome to 5,6-dihydroxyindole (DHI) and 5,6-dihydroxyindole-2-carboxylic acid (DHICA), and the rearrangement of 5-S-cysteinyldopa o-quinoneimine to 1,4-benzothiazine (BTZ) and its 3-carboxylic acid (BTZCA). In eumelanins, the DHICA-to-DHI ratio markedly affects the overall antioxidant and paramagnetic properties of the resulting pigments. In particular, a higher content in DHICA decreases visible light absorption and paramagnetic response relative to DHI-based melanins, but markedly enhances antioxidant properties. In pheomelanins, likewise, BTZCA-related units, prevalently formed in the presence of zinc ions, appear to confer pronounced visible and ultraviolet A (UVA) absorption features, accounting for light-dependent reactive oxygen species (ROS) production, whereas non-carboxylated benzothiazine intermediates seem to be more effective in inducing ROS production by redox cycling mechanisms in the dark. The possible biological and functional significance of carboxyl retention in the eumelanin and pheomelanin pathways is discussed.

Keywords: 5,6-dihydroxyindoles; 5-S-cysteinyldopa; antioxidant; benzothiazines; dopachrome; eumelanin; melanins; melanocortin-1-receptor; pheomelanin; pro-oxidant.

Scheme 1

Schematic view of eumelanin and pheomelanin biosynthetic pathways.

Figure 1

Main oxidation products of DHI (blue box) and DHICA (yellow box).

Figure 2

Predicted structures and experimental UV-visible absorption spectra at various times of the oxidation mixtures of DHI and DHICA oligomers. Blue box: change in absorption at various times after pulse radiolysis of a N₂O-satured aqueous solution of DHI dimer (1.5 × 10⁻⁴ M) in 0.5 M KBr/7.0 × 10⁻² M phosphate buffer, pH 7.0: (○) 14 μs; (■) 102 μs; (▲) 402 μs; (♦) 866 μs. Yellow box: changes in absorption at various times after pulse radiolysis of DHICA dimer (1.5 × 10⁻⁴ M) in 0.5 M KBr/7.0 × 10⁻² phosphate buffer, pH 7.0: (○) 400 μs; (■) 5 μs; (Δ) 44 ms [31,32].

Figure 3

UV-visible spectra of DHI (black trace) and DHICA melanin (gray trace) at pH 7.5.

Figure 4

Melanins from DHI (blue box) and DHICA (yellow box) oxidation before and after 1:90 dilution in 1% PVA-containing buffer.

Figure 5

Intrinsic and extrinsic contributions to the chromophoric properties of DHI and DHICA melanin.

Figure 6

Simplified representation of aggregates’ formation for DHI and DHICA melanins.

Figure 7

Main oxidation products of BTZ (blue box) and BTZCA (yellow box).

Figure 8

Glutathione (GSH) depletion (%) after 3 h in 0.1 M phosphate buffer (pH 7.4) in the absence or in the presence of 5SCD melanin prepared in the absence or in the presence of Zn²⁺ ions. Reported are the mean ± SD values from at least five experiments. Significant differences were determined by an independent sample two-tailed t-test.