Effect of cyano-addition on the photoacidity switch in 5-cyano-8-amino-2-naphthol.

Cyano- or CN-additions are often utilized in the design of photoacids to enhance and/or enable excited state proton transfer (ESPT) from the protic site to aqueous and nonaqueous solvents. In diprotic photoacid 8-amino-2-naphthol (8N2OH), the protonation state of the amino group (NH3+/NH2) acts as an on-off switch for ESPT at the OH site in water. This study investigated whether the addition of CN in 5-cyano-8-amino-2-naphthol (5CN8) could override this switch and promote new ESPT pathways. Analysis of the steady-state and time-resolved emission data showed that in the presence of protonated NH3+, CN enhances OH photoacidity (vs. in 8N2OH) and activates the ESPT pathway at NH3+. Both protic sites, OH and NH3+, can also donate a proton to methanol upon excitation. In contrast, in the presence of deprotonated NH2, despite the addition of CN, ESPT is still not observed at the OH site for 5CN8. Thus, the addition of CN cannot override or negate the inhibiting effect of NH2 on OH photoacidity. Potential causes for this inhibition are discussed, including electronic and antiaromaticity effects of CN and NH2 substitution.

Photophysical properties of a boron analogue of coumarin.

In this report, we present a study into the structure and electronic properties of difluoroboronsalicylaldoxime (DFBS), a boron-based structural analog of coumarin. The modification of the heterocyclic ring of coumarin with boron results in a compound with similar structural parameters and molecular orbitals to coumarin. DFT and TDDFT calculations reveal a significant stabilization of the LUMO in DFBS; this is supported by a ∼40 nm red shift of the lowest electronic transition in the absorption spectrum. Interestingly, DFBS is emissive, while unmodified coumarin is effectively non-radiative. Comparisons between DFBS, emissive coumarin variants, and unmodified coumarin suggest that the charge transfer character of the transition contributes to the fluorescence.

Light-Induced Nanosecond Relaxation Dynamics of Rhenium-Labeled Pseudomonas aeruginosa Azurins.

Time-resolved phosphorescence spectra of Re(CO)3(dmp)+ and Re(CO)3(phen)+ chromophores (dmp = 4,7-dimethyl-1,10-phenanthroline, phen = 1,10-phenanthroline) bound to surface histidines (H83, H124, and H126) of Pseudomonas aeruginosa azurin mutants exhibit dynamic band maxima shifts to lower wavenumbers following 3-exponential kinetics with 1-5 and 20-100 ns major phases and a 1.1-2.5 µs minor (5-16%) phase. Observation of slow relaxation components was made possible by using an organometallic Re chromophore as a probe whose long phosphorescence lifetime extends the observation window up to ∼3 µs. Integrated emission-band areas also decay with 2- or 3-exponential kinetics; the faster decay phase(s) is relaxation-related, whereas the slowest one [360-680 ns (dmp); 90-140 ns (phen)] arises mainly from population decay. As a result of shifting bands, the emission intensity decay kinetics depend on the detection wavelength. Detailed kinetics analyses and comparisons with band-shift dynamics are needed to disentangle relaxation and population decay kinetics if they occur on comparable timescales. The dynamic phosphorescence Stokes shift in Re-azurins is caused by relaxation motions of the solvent, the protein, and solvated amino acid side chains at the Re binding site in response to chromophore electronic excitation. Comparing relaxation and decay kinetics of Re(dmp)124K122CuII and Re(dmp)124W122CuII suggests that electron transfer (ET) and relaxation motions in the W122 mutant are coupled. It follows that nanosecond and faster photo-induced ET steps in azurins (and likely other redox proteins) occur from unrelaxed systems; importantly, these reactions can be driven (or hindered) by structural and solvational dynamics.

Divergent excited state proton transfer reactions of bifunctional photoacids 1-ammonium-2-naphthol and 3-ammonium-2-naphthol in water and methanol.

This paper highlights the challenge of predicting the excited state proton transfer (ESPT) reactions of small organic compounds with multiple proton transfer sites. Aminonaphthols, naphthalene compounds with both hydroxyl and amino substituents, can be viewed as a combination of two monoprotic photoacids, naphthol and naphthylammonium. Here, the ESPT reactions of 3-ammonium-2-naphthol (3N2OH) and 1-ammonium-2-naphthol (1N2OH) were studied in water and methanol using a combination of steady-state and time-correlated single-photon counting emission spectroscopy. For 3N2OH, ESPT was observed at the OH site in water but at neither of the sites in methanol; for 1N2OH, ESPT was observed at both the OH and NH3+ sites in water but only at the NH3+ site in methanol. Evidence of ESPT at the NH3+ site is limited for aminonaphthols. The divergent dynamics of 3N2OH and 1N2OH in water and methanol are discussed; dependent on the substitution and solvent, the ESPT reactions were analysed within the frameworks of reference photoacids 2-naphthol and 1-naphthylammonium. The application of crown ether and salt to control the release of select protons in non-aqueous media is also discussed.

Divergent Hammett Plots of the Ground- and Excited-State Proton Transfer Reactions of 7-Substituted-2-Naphthol Compounds.

The rational design of photoacids requires accessible predictive models of the electronic effect of functional groups on chemical templates of interest. Here, the effect of substituents on the photoacidity and excited-state proton transfer (PT) pathways of prototype 2-naphthol (2OH) at the symmetric C7 position was investigated through photochemical and computational studies of 7-amino-2-naphthol (7N2OH) and 7-methoxy-2-naphthol (7OMe2OH). Time-resolved emission experiments of 7N2OH revealed that the presence of an electron-withdrawing versus electron-donating group (EWG vs EDG, NH3+ vs NH2) led to a drastic decline in photoacidity: p Ka* = 1.1 ± 0.2 vs 9.6 ± 0.2. Time-dependent density functional theory calculations with explicit water molecules confirmed that the excited neutral state (x = NH2) is greatly stabilized by water, with equation-of-motion coupled cluster singles and doubles calculations supporting potential mixing between the La and Lb states. Similar suppression of photoacidity, however, was not observed for 7OMe2OH with EDG OCH3, p Ka* = 2.7 ± 0.1. Hammett plots of the ground- and excited-state PT reactions of substituted 7-x-2OH compounds (x = CN, NH3+, H, CH3, OCH3, OH, and NH2) vs Hammett parameters σp showed breaks in the linearity between the EDG and EWG regions: ρ ∼ 0 vs 1.14 and ρ* ∼ 0 vs 3.86. The divergent acidic behavior most likely arises from different mixing mechanisms of the lowest Lb state with the La and possible Bb states upon substitution of naphthalene in water.

Two Tryptophans Are Better Than One in Accelerating Electron Flow through a Protein.

We have constructed and structurally characterized a Pseudomonas aeruginosa azurin mutant Re126WWCuI , where two adjacent tryptophan residues (W124 and W122, indole separation 3.6-4.1 Å) are inserted between the CuI center and a Re photosensitizer coordinated to the imidazole of H126 (ReI(H126)(CO)3(4,7-dimethyl-1,10-phenanthroline)+). CuI oxidation by the photoexcited Re label (*Re) 22.9 Å away proceeds with a ∼70 ns time constant, similar to that of a single-tryptophan mutant (∼40 ns) with a 19.4 Å Re-Cu distance. Time-resolved spectroscopy (luminescence, visible and IR absorption) revealed two rapid reversible electron transfer steps, W124 → *Re (400-475 ps, K 1 ≅ 3.5-4) and W122 → W124•+ (7-9 ns, K 2 ≅ 0.55-0.75), followed by a rate-determining (70-90 ns) CuI oxidation by W122•+ ca. 11 Å away. The photocycle is completed by 120 µs recombination. No photochemical CuI oxidation was observed in Re126FWCuI , whereas in Re126WFCuI , the photocycle is restricted to the ReH126W124 unit and CuI remains isolated. QM/MM/MD simulations of Re126WWCuI indicate that indole solvation changes through the hopping process and W124 → *Re electron transfer is accompanied by water fluctuations that tighten W124 solvation. Our finding that multistep tunneling (hopping) confers a ∼9000-fold advantage over single-step tunneling in the double-tryptophan protein supports the proposal that hole-hopping through tryptophan/tyrosine chains protects enzymes from oxidative damage.

Hole Hopping Across a Protein-Protein Interface.

We have investigated photoinduced hole hopping in a Pseudomonas aeruginosa azurin mutant Re126WWCuI, where two adjacent tryptophan residues (W124 and W122) are inserted between the CuI center and a Re photosensitizer coordinated to a H126 imidazole (Re = ReI(H126)(CO)3(dmp)+, dmp = 4,7-dimethyl-1,10-phenanthroline). Optical excitation of this mutant in aqueous media (≤40 µM) triggers 70 ns electron transport over 23 Å, yielding a long-lived (120 µs) ReI(H126)(CO)3(dmp•-)WWCuII product. The Re126FWCuI mutant (F124, W122) is not redox-active under these conditions. Upon increasing the concentration to 0.2-2 mM, {Re126WWCuI}2 and {Re126FWCuI}2 are formed with the dmp ligand of the Re photooxidant of one molecule in close contact (3.8 Å) with the W122' indole on the neighboring chain. In addition, {Re126WWCuI}2 contains an interfacial tryptophan quadruplex of four indoles (3.3-3.7 Å apart). In both mutants, dimerization opens an intermolecular W122' → //*Re ET channel (// denotes the protein interface, *Re is the optically excited sensitizer). Excited-state relaxation and ET occur together in two steps (time constants of ∼600 ps and ∼8 ns) that lead to a charge-separated state containing a Re(H126)(CO)3(dmp•-)//(W122•+)' unit; then (CuI)' is oxidized intramolecularly (60-90 ns) by (W122•+)', forming ReI(H126)(CO)3(dmp•-)WWCuI//(CuII)'. The photocycle is closed by ∼1.6 µs ReI(H126)(CO)3(dmp•-) → //(CuII)' back ET that occurs over 12 Å, in contrast to the 23 Å, 120 µs step in Re126WWCuI. Importantly, dimerization makes Re126FWCuI photoreactive and, as in the case of {Re126WWCuI}2, channels the photoproduced "hole" to the molecule that was not initially photoexcited, thereby shortening the lifetime of ReI(H126)(CO)3(dmp•-)//CuII. Although two adjacent W124 and W122 indoles dramatically enhance CuI → *Re intramolecular multistep ET, the tryptophan quadruplex in {Re126WWCuI}2 does not accelerate intermolecular electron transport; instead, it acts as a hole storage and crossover unit between inter- and intramolecular ET pathways. Irradiation of {Re126WWCuII}2 or {Re126FWCuII}2 also triggers intermolecular W122' → //*Re ET, and the Re(H126)(CO)3(dmp•-)//(W122•+)' charge-separated state decays to the ground state by ∼50 ns ReI(H126)(CO)3(dmp•-)+ → //(W122•+)' intermolecular charge recombination. Our findings shed light on the factors that control interfacial hole/electron hopping in protein complexes and on the role of aromatic amino acids in accelerating long-range electron transport.

pH switch for OH-photoacidity in 5-amino-2-naphthol and 8-amino-2-naphthol.

Photoactive charge transfer compounds are of strong interest for their potential applications in material, chemical, and biological science and their abilities to elucidate fundamental charge transfer mechanisms. Aminonaphthols, photoacids with both oxygen (OH) and nitrogen-based (NH2) protonation sites, have been reported to undergo simultaneous excited-state proton transfer (ESPT) in water upon excitation. In this paper, the ESPT mechanism for zwitterion formation in 8-amino-2-naphthol (8N2OH) and 5-amino-2-naphthol (5N2OH) was examined using a combination of time-resolved emission spectroscopy and time-dependent density functional theory (TD-DFT) calculations. The measurements prompted a re-assignment of the zwitterion state in the steady-state emission spectra; analysis of the time-correlated single-photon counting emission data showed that the zwitterion was formed only from excitation of protonated 5N2OH and 8N2OH such that ESPT occurred only at the single hydroxyl group. The protonation state of the amino group dramatically altered the photoacidity of OH, such that the pH behaved as an on/off switch for photoacidity. In the protonated state (NH3+), the pKa*(OH) values of 5N2OH and 8N2OH were both 1.1 ± 0.2, while in the deprotonated state (NH2), the two pKa*(OH) values were similar to the ground state proton acidity, pKa(OH) = 9.5 ± 0.2. The switching of the photoacidity was investigated using TD-DFT calculations and the linear free energy Hammett relation. The latter was shown to not describe the excited state data over the broad pH range.

Two-photon spectroscopy of tungsten(0) arylisocyanides using nanosecond-pulsed excitation.

The two-photon absorption (TPA) cross sections (δ) for tungsten(0) arylisocyanides (W(CNAr)6) were determined in the 800-1000 nm region using two-photon luminescence (TPL) spectroscopy. The complexes have high TPA cross sections, in the range 1000-2000 GM at 811.8 nm. In comparison, the cross section at 811.8 nm for tris-(2,2'-bipyridine)ruthenium(ii), [Ru(bpy)3]2+, is 7 GM. All measurements were performed using a nanosecond-pulsed laser system.

Tryptophan-accelerated electron flow across a protein-protein interface.

We report a new metallolabeled blue copper protein, Re126W122Cu(I) Pseudomonas aeruginosa azurin, which has three redox sites at well-defined distances in the protein fold: Re(I)(CO)3(4,7-dimethyl-1,10-phenanthroline) covalently bound at H126, a Cu center, and an indole side chain W122 situated between the Re and Cu sites (Re-W122(indole) = 13.1 Å, dmp-W122(indole) = 10.0 Å, Re-Cu = 25.6 Å). Near-UV excitation of the Re chromophore leads to prompt Cu(I) oxidation (<50 ns), followed by slow back ET to regenerate Cu(I) and ground-state Re(I) with biexponential kinetics, 220 ns and 6 µs. From spectroscopic measurements of kinetics and relative ET yields at different concentrations, it is likely that the photoinduced ET reactions occur in protein dimers, (Re126W122Cu(I))2 and that the forward ET is accelerated by intermolecular electron hopping through the interfacial tryptophan: *Re//←W122←Cu(I), where // denotes a protein-protein interface. Solution mass spectrometry confirms a broad oligomer distribution with prevalent monomers and dimers, and the crystal structure of the Cu(II) form shows two Re126W122Cu(II) molecules oriented such that redox cofactors Re(dmp) and W122-indole on different protein molecules are located at the interface at much shorter intermolecular distances (Re-W122(indole) = 6.9 Å, dmp-W122(indole) = 3.5 Å, and Re-Cu = 14.0 Å) than within single protein folds. Whereas forward ET is accelerated by hopping through W122, BET is retarded by a space jump at the interface that lacks specific interactions or water molecules. These findings on interfacial electron hopping in (Re126W122Cu(I))2 shed new light on optimal redox-unit placements required for functional long-range charge separation in protein complexes.