Background: Melanin is a poorly understood fungal virulence factor. Results: 2D 13 C-13 C correlation solid-state nuclear magnetic resonance reveals the carbon-based molecular architecture of intact melanin pigment assemblies in Cryptococcus neoformans. Conclusion: Polysaccharide cell-wall components form a scaffold for layered deposition of aromatic-based pigment assemblies. Significance: Deciphering macromolecular interactions that drive melanin pigment assembly in fungal cell walls facilitates the development of drug delivery materials.

Melanins are a class of natural pigments associated with a wide range of biological functions, including microbial virulence, energy transduction, and protection against solar radiation. Because of their insolubility and structural heterogeneity, solid-state nuclear magnetic resonance (NMR) spectroscopy provides an unprecedented means to define the molecular architecture of these enigmatic pigments. The requirement of obligatory catecholamines for melanization of the pathogenic fungus Cryptococcus neoformans also offers unique opportunities for investigating melanin development. In the current study, pigments produced with L-dopa, methyl-L-dopa, epinephrine, and norepinephrine precursors are compared structurally using 13 C and 1 H magic-angle spinning (MAS) NMR. Striking structural differences were observed for both aromatic and aliphatic molecular constituents of the mature fungal pigment assemblies, thus making it possible to redefine the molecular prerequisites for formation of the aromatic domains of insoluble indole-based biopolymers, to rationalize their distinctive physical characteristics, and to delineate the role of cellular constituents in assembly of the melanized macromolecules with polysaccharides and fatty acyl chain-containing moieties. By achieving an augmented understanding of the mechanisms of C. neoformans melanin biosynthesis and cellular assembly, such studies can guide future drug discovery efforts related to melaninassociated virulence, resistance to tumor therapy, and production of melanin mimetics under cell-free conditions.

An experimental strategy has been developed to increase the efficiency of dynamic nuclear polarization (DNP) in solid-state NMR studies. The method makes assignments simpler, faster, and more reliable via sequential correlations of both side-chain and Cα resonances. The approach is particularly suited to complex biomolecules and systems with significant chemical-shift degeneracy. It was designed to overcome the spectral congestion and line broadening that occur due to sample freezing at the cryogenic temperatures required for DNP. Nonuniform sampling (NUS) is incorporated to achieve time-efficient collection of multidimensional data. Additionally, fast (25 kHz) magic-angle spinning (MAS) provides optimal sensitivity and resolution. Data collected in <1 wk produced a virtually complete de novo assignment of the coat protein of Pf1 virus. The peak positions and linewidths for samples near 100 K are perturbed relative to those near 273 K. These temperature-induced perturbations are strongly correlated with hydration surfaces.

Graphene nanoribbons (GNRs) with robust electronic band gaps are promising candidate materials for nanometer-scale electronic circuits. Realizing their full potential, however, will depend on the ability to access GNRs with prescribed widths and edge structures and an understanding of their fundamental electronic properties. We report field-effect devices exhibiting ambipolar transport in accumulation mode composed of solution-synthesized GNRs with straight armchair edges. Temperature-dependent electrical measurements specify thermally activated charge transport, which we attribute to inter-ribbon hopping. With access to structurally precise materials in practical quantities and by overcoming processing difficulties in making electrical contacts to these materials, we have demonstrated critical steps toward nanoelectric devices based on solution-synthesized GNRs.

Despite the essential functions of melanin pigments in diverse organisms and their roles in inspiring designed nanomaterials for electron transport and drug delivery, the structural frameworks of the natural materials and their biomimetic analogs remain poorly understood. To overcome the investigative challenges posed by these insoluble heterogeneous pigments, we have used L-tyrosine or dopamine enriched with stable 13C and 15N isotopes to label eumelanins metabolically in cell-free and Cryptococcus neoformans cell systems and to define their molecular structures and supramolecular architectures. Using high-field two-dimensional solid-state nuclear magnetic resonance (NMR), our study directly evaluates the assumption of structural commonality between synthetic melanin models and the corresponding natural pigments, demonstrating a common indole-based aromatic core in the products from contrasting synthetic protocols for the first time.