Photochemical Tyrosine Oxidation with a Hydrogen-Bonded Proton Acceptor by Bidirectional Proton-Coupled Electron Transfer.
ABSTRACT: Amino acid radical generation and transport are fundamentally important to numerous essential biological processes to which small molecule models lend valuable mechanistic insights. Pyridyl-amino acid-methyl esters are appended to a rhenium(I) tricarbonyl 1,10-phenanthroline core to yield rhenium-amino acid complexes with tyrosine ([Re]-Y-OH) and phenylalanine ([Re]-F). The emission from the [Re] center is more significantly quenched for [Re]-Y-OH upon addition of base. Time-resolved studies establish that excited-state quenching occurs by a combination of static and dynamic mechanisms. The degree of quenching depends on the strength of the base, consistent with a proton-coupled electron transfer (PCET) quenching mechanism. Comparative studies of [Re]-Y-OH and [Re]-F enable a detailed mechanistic analysis of a bidirectional PCET process.
Project description:Rhenium-based anticancer agents have arisen as promising alternatives to conventional platinum-based drugs. Based on previous studies demonstrating how increasing lipophilicity improves drug uptake within the cell, we sought to investigate the effects of lipophilicity on the anticancer activity of a series of six rhenium(i) tricarbonyl complexes. These six rhenium(i) tricarbonyl structures, called Re-Chains, bear pyridyl imine ligands with different alkyl chains ranging in length from two to twelve carbons. The cytotoxicities of these compounds were measured in HeLa cells. At long timepoints (48 h), all compounds are equally cytotoxic. At shorter time points, however, the compounds with longer alkyl chains are significantly more active than those with smaller chains. Cellular uptake studies of these compounds show that they are taken up via both passive and active pathways. Collectively, these studies show how lipophilicity affects the rate at which these Re compounds induce their biological activities.
Project description:Photochemical radical initiation is a powerful tool for studying radical initiation and transport in biology. Ribonucleotide reductases (RNRs), which catalyze the conversion of nucleotides to deoxynucleotides in all organisms, are an exemplar of radical mediated transformations in biology. Class Ia RNRs are composed of two subunits: ?2 and ?2. As a method to initiate radical formation photochemically within ?2, a single surface-exposed cysteine of the ?2 subunit of Escherichia coli Class Ia RNR has been labeled (98%) with a photooxidant ([Re ] = tricarbonyl(1,10-phenanthroline)(methylpyridyl)rhenium(I)). The labeling was achieved by incubation of S355C-?2 with the 4-(bromomethyl)pyridyl derivative of [Re] to yield the labeled species, [Re]-S355C-?2. Steady-state and time-resolved emission experiments reveal that the metal-to-ligand charge transfer (MLCT) excited-state (3)[Re ](?) is not significantly perturbed after bioconjugation and is available as a phototrigger of tyrosine radical at position 356 in the ?2 subunit; transient absorption spectroscopy reveals that the radical lives for microseconds. The work described herein provides a platform for photochemical radical initiation and study of proton-coupled electron transfer (PCET) in the ?2 subunit of RNR, from which radical initiation and transport for this enzyme originates.
Project description:Proton-coupled electron transfer (PCET) underpins energy conversion in biology. PCET may occur with the unidirectional or bidirectional transfer of a proton and electron and may proceed synchronously or asynchronously. To illustrate the role of PCET in biology, this review presents complementary biological and model systems that explore PCET in electron transfer (ET) through hydrogen bonds [azurin as compared to donor-acceptor (D-A) hydrogen-bonded networks], the activation of C-H bonds [alcohol dehydrogenase and soybean lipoxygenase (SLO) as compared to Fe(III) metal complexes], and the generation and transport of amino acid radicals [photosystem II (PSII) and ribonucleotide reductase (RNR) as compared to tyrosine-modified photoactive Re(I) and Ru(II) complexes]. In providing these comparisons, the fundamental principles of PCET in biology are illustrated in a tangible way.
Project description:The reaction of Re(CO)(3)(H(2)O)(3)(+) with hen egg lysozyme in aqueous solution results in a single covalent adduct; single crystal X-ray diffraction shows that the rhenium tricarbonyl cation binds to His15 in two significantly populated rotamer conformations.
Project description:Aromatic amino acids such as l-tyrosine and l-tryptophan are deployed in natural systems to mediate electron transfer (ET) reactions. While tyrosine oxidation is always coupled to deprotonation (proton-coupled electron-transfer, PCET), both ET-only and PCET pathways can occur in the case of the tryptophan residue. In the present work, two novel conjugates 1 and 2, based on a Sn<sup>IV</sup> tetraphenylporphyrin and Sn<sup>IV</sup> octaethylporphyrin, respectively, as the chromophore/electron acceptor and l-tryptophan as electron/proton donor, have been prepared and thoroughly characterized by a combination of different techniques including single crystal X-ray analysis. The photophysical investigation of 1 and 2 in CH<sub>2</sub> Cl<sub>2</sub> in the presence of pyrrolidine as a base shows that different quenching mechanisms are operating upon visible-light excitation of the porphyrin component, namely photoinduced electron transfer and concerted proton electron transfer (CPET), depending on the chromophore identity and spin multiplicity of the excited state. The results are compared with those previously described for metal-mediated analogues featuring Sn<sup>IV</sup> porphyrin chromophores and l-tyrosine as the redox active amino acid and well illustrate the peculiar role of l-tryptophan with respect to PCET.
Project description:The reaction of rhenium α-diimine (N-N) tricarbonyl complexes with nitrosonium tetrafluoroborate yields the corresponding dicarbonyl-nitrosyl [Re(CO)<sub>2</sub>(NO)(N-N)X]<sup>+</sup> species (where X = halide). The complexes, accessible in a single step in good yield, are structurally nearly identical higher charge congeners of the tricarbonyl molecules. Substitution chemistry aimed at the realization of equivalent dicationic species (intended for applications as potential antimicrobial agents), revealed that the reactivity of metal ion in [Re(CO)<sub>2</sub>(NO)(N-N)X]<sup>+</sup> is that of a hard Re acid, probably due to the stronger π-acceptor properties of NO<sup>+</sup> as compared to those of CO. The metal ion thus shows great affinity for π-basic ligands, which are consequently difficult to replace by, e.g., σ-donor or weak π-acids like pyridine. Attempts of direct nitrosylation of α-diimine <i>fac</i>-[Re(CO)<sub>3</sub>]<sup>+</sup> complexes bearing π-basic OR-type ligands gave the [Re(CO)<sub>2</sub>(NO)(N-N)(BF<sub>4</sub>)][BF<sub>4</sub>] salt as the only product in good yield, featuring a stable Re-FBF<sub>3</sub> bond. The solid state crystal structure of nearly all molecules presented could be elucidated. A fundamental consequence of the chemistry of [Re(CO)<sub>2</sub>(NO)(N-N)X]<sup>+</sup> complexes, it that the same can be photo-activated towards CO release and represent an entirely new class of photoCORMs.
Project description:The synthesis and full characterization of two tetrathiafulvalene-appended azine ligands, namely 2-([2,2'-bi(1,3-dithiolylidene)]-4-yl)-6-((2,4-dinitrophenyl)hydrazono)methyl)pyridine (L1) and 5-([2,2'-bi(1,3-dithiolylidene)]-4-yl)-2-((2,4-dinitrophenyl)hydrazono)methyl)pyridine (L2) are described. The crystal structure of ligand L1 indicates that the ligand is completely planar with the presence of a strong intramolecular N3-H3···O1 hydrogen bonding. Titration experiments with inorganic anions showed that both ligands are suitable candidates for the sensing of fluoride anions. Ligand L2 was reacted with a Re(I) cation to yield the corresponding rhenium tricarbonyl complex 3. In the crystal structure of the newly prepared electroactive rhenium complex the TTF is neutral and the rhenium cation is hexacoordinated. The electrochemical behavior of the three compounds indicates that they are promising for the construction of crystalline radical cation salts.
Project description:As part of our program to develop new probes for the estrogen receptor binding domain, we prepared and evaluated a novel 17?-(rhenium tricarbonyl bipyridyl) vinyl estradiol complex. Preparation of the final compound was achieved using the Stille coupling between the preformed brominated rhenium tricarbonyl bipyridine complex and the tributylstannyl vinyl estradiol. Competitive receptor binding assays and stimulatory assays demonstrated that the final complex retained affinity and efficacy comparable to the corresponding pyridyl vinyl estradiol analog, but lower than that of the phenyl vinyl estradiol analog.
Project description:The use of light to drive proton-coupled electron transfer (PCET) reactions has received growing interest, with recent focus on the direct use of excited states in PCET reactions (ES-PCET). Electrostatic ion pairs provide a scaffold to reduce reaction orders and have facilitated many discoveries in electron-transfer chemistry. Their use, however, has not translated to PCET. Herein, we show that ion pairs, formed solely through electrostatic interactions, provide a general, facile means to study an ES-PCET mechanism. These ion pairs formed readily between salicylate anions and tetracationic ruthenium complexes in acetonitrile solution. Upon light excitation, quenching of the ruthenium excited state occurred through ES-PCET oxidation of salicylate within the ion pair. Transient absorption spectroscopy identified the reduced ruthenium complex and oxidized salicylate radical as the primary photoproducts of this reaction. The reduced reaction order due to ion pairing allowed the first-order PCET rate constants to be directly measured through nanosecond photoluminescence spectroscopy. These PCET rate constants saturated at larger driving forces consistent with approaching the Marcus barrierless region. Surprisingly, a proton-transfer tautomer of salicylate, with the proton localized on the carboxylate functional group, was present in acetonitrile. A pre-equilibrium model based on this tautomerization provided non-adiabatic electron-transfer rate constants that were well described by Marcus theory. Electrostatic ion pairs were critical to our ability to investigate this PCET mechanism without the need to covalently link the donor and acceptor or introduce specific hydrogen bonding sites that could compete in alternate PCET pathways.
Project description:Rhenium tricarbonyl complexes have been recently investigated as novel anticancer agents. However, little is understood about their mechanisms of action, as well as the means by which cancer cells respond to chronic exposure to these compounds. To gain a deeper mechanistic insight into these rhenium anticancer agents, we developed and characterized an ovarian cancer cell line that is resistant to a previously studied compound [Re(CO)<sub>3</sub> (dmphen)(ptolICN)]<sup>+</sup> , where dmphen=2,9-dimethyl-1,10-phenanthroline and ptolICN=para-tolyl isonitrile, called TRIP. This TRIP-resistant ovarian cancer cell line, A2780TR, was found to be 9 times less sensitive to TRIP compared to the wild-type A2780 ovarian cancer cell line. Furthermore, the cytotoxicities of established drugs and other rhenium anticancer agents in the TRIP-resistant cell line were determined. Notably, the drug taxol was found to exhibit a 184-fold decrease in activity in the A2780TR cell line, suggesting that mechanisms of resistance towards TRIP and this drug are similar. Accordingly, expression levels of the ATP-binding cassette transporter P-glycoprotein, an efflux transporter known to detoxify taxol, were found to be elevated in the A2780TR cell line. Additionally, a gene expression analysis using the National Cancer Institute 60 cell line panel identified the MT1E gene to be overexpressed in cells that are less sensitive to TRIP. Because this gene encodes for metallothioneins, this result suggests that detoxification by this class of proteins is another mechanism for resistance to TRIP. The importance of this gene in the A2780TR cell line was assessed, confirming that its expression is elevated in this cell line as well. As the first study to investigate and identify the cancer cell resistance pathways in response to a rhenium complex, this report highlights important similarities and differences in the resistance responses of ovarian cancer cells to TRIP and conventional drugs.