Sequence-dependent interactions of cationic naphthalimides and polynucleotides.

The binding interactions of three naphthalimide derivatives with heteropoly nucleic acids have been evaluated using fluorescence, absorption and circular dichroism spectroscopies. Mono- and bifunctionalized naphthalimides exhibit sequence-dependent variations in their affinity toward DNA. The heteropoly nucleic acids, [Poly(dA-dT)]2 and [Poly(dG-dC)]2, as well as calf thymus (CT) DNA, were used to understand the factors that govern binding strength and selectivity. Sequence selectivity was addressed by determining the binding constants as a function of polynucleotide composition according to the noncooperative McGhee-von Hippel binding model. Binding affinities toward [poly(dA-dT)](2) were the largest for spermine-substituted naphthalimides (Kb = 2-6 x 10(6) M(-1)). The association constants for complex formation between the cationic naphthalimides and [poly(dG-dC)]2 or CT DNA (58% A-T content) were 2-500 times smaller, depending on the naphthalimide-polynucleotide pair. The binding modes were also assessed using a combination of induced circular dichroism and salt effects to determine whether the naphthalimides associate with DNA through intercalative, electrostatic or groove-binding. The results show that the monofunctionalized spermine and pyridinium-substituted naphthalimides associate with DNA through electrostatic interactions. In contrast, intercalative interactions are predominant in the complex formed between the bifunctionalized spermine compound and all of the polynucleotides.

Effect of electron mediators on current generation and fermentation in a microbial fuel cell.

Effects of select electron mediators [9,10-anthraquinone-2,6-disulfonic acid disodium salt (AQDS), safranine O, resazurin, methylene blue, and humic acids] on metabolic end-products and current production from cellulose digestion by Clostridium cellulolyticum in microbial fuel cells (MFCs) were studied using capillary electrophoresis and traditional electrochemical techniques. Addition of the mediator resazurin greatly enhanced current production but did not appear to alter the examined fermentation end-products compared to MFCs with no mediator. Assays for lactate, acetate, and ethanol indicate that the presence of safranine O, methylene blue, and humic acids alters metabolite production in the MFC: safranine O decreased the examined metabolites, methylene blue increased lactate formation, and humic acids increased the examined metabolites. Mediator standard redox potentials (E (0)) reported in the literature do not coincide with redox potentials in MFCs due presumably to the electrolytic complexity of media that supports bacterial survival and growth. Current production in MFCs: (1) can be effected by the mediator redox potential while in the media, which may be significantly shifted from E (0), and (2) depended on the ability of the mediator to access the bacterial electron source, which may be cytoplasmic. In addition, some electron mediators had significant effects on metabolic end-products and therefore the metabolism of the organism itself.

Ground-state interactions of spermine-substituted naphthalimides with mononucleotides.

Water soluble spermine, spermine-naphthalimide, and pyridinium-substituted 1,8-naphthalimide derivatives have been synthesized as nucleotide-specific binding agents. Both mono- and bifunctionalized spermine compounds were studied. The photophysical properties of each compound were studied by using time-resolved and steady-state fluorescence and absorption spectroscopies. The fluorescence decay of the mononaphthalimides was adequately fit to a single exponential decay, and in all cases, the lifetime (2.4 ns) was independent of the imide substitutent. In the case of the bisnaphthalimide, emission from both the monomer and ground-state dimer forms was observed. The fluorescence quantum yield of the monomer (0.03) was significantly smaller than that of the mononaphthalimides (0.27). The dimer emission was red-shifted relative to that of the monomer. The singlet-state lifetime of the dimer was found to be 20 ns. In all cases, only absorption from the triplet excited state was observed, indicating no evidence of a naphthalimide radical anion from dimer excitation. The ground-state interactions of the naphthalimides with four nucleotides were investigated. Nucleotide selectivity was evaluated by determining their individual binding constants (Keq). The association constants were measured by using absorption, time-resolved fluorescence, and combined time-resolved and steady-state fluorescence. The equilibrium binding constant was largest for association of the spermine-substituted mononaphthalimide with adenosine monophosphate (Keq=550 M-1) or guanosine monophosphate (Keq=440 M-1). The dimer form of the disubstituted spermine also showed binding constants in excess of 200 M-1 with the purine nucleotides. The association constant for binding of the pyridinium-substituted naphthalimide showed little dependence on the structure of the nucleotide.

Reactivities of carboxyalkyl-substituted 1,4,5,8-naphthalene diimides in aqueous solution.

A series of water-soluble 1,4,5,8-naphthalene diimide derivatives has been prepared and their redox and photophysical properties characterized. From laser flash photolysis studies, the triplet excited state of N,N'-bis[2-(N-pyridinium)ethyl]-1,4,5,8-naphthalene diimide (NDI-pyr) was found to undergo oxidative quenching with the electron donors DABCO, tyrosine, and tryptophan as expected from thermodynamics. Interestingly, the reactivities of naphthalene diimides (NDI) possessing alpha- and beta-carboxylic acid substituents (R = -CH2COO-, -C(CH3)2COO-, and -CH2CH2COO-) were strikingly different. In these compounds, the transient produced upon 355 nm excitation did not react with the electron donors. Instead, this transient reacted rapidly (k > 10(8)-10(9) M-1 s-1) with known electron acceptors, benzyl viologen and ferricyanide. The transient spectrum of the carboxyalkyl-substituted naphthalimides observed immediately after the laser pulse was nearly identical to the one-electron-reduced form of 1,4,5,8-naphthalene diimide (produced independently using the bis-pyridinium-substituted naphthaldiimide). From our studies, we conclude that the transient produced upon nanosecond laser flash photolysis of NDI-(CH2)nCOO- is the species produced upon intramolecular electron transfer from the carboxylate moiety to the singlet excited state of NDI. In separate experiments, we verified that the singlet excited state of NDI-pyr does, indeed, react intermolecularly with acetate, alanine, and glycine. The process is further substantiated using thermodynamic driving force calculations. The results offer new prospects of the efficient photochemical production of reactive carbon-centered radicals.