Sameh H. Abdelwahed

Crossover from Single-Step Tunneling to Multistep Hopping for Molecular Triplet Energy Transfer

Tracking Triplet-State Transfers In devices such as organic transistors and photovoltaic cells, energy flow from donor to acceptor sites can occur via electrons that have been excited into higher electronic levels, which create a triplet state with two unpaired spins. At short distances between donor and acceptor, the transfer occurs through direct tunneling, but at longer distances, the electron “hops” in a multistep process. Vura-Weis et al. (p. 1547) used femtosecond transient absorption spectroscopy to observe this crossover in transfer mechanism directly in a series of molecules with varying bridge lengths between the donor and acceptor. Ultrafast absorption spectroscopy reveals a change in mechanism when sites for electron transfer become more distant. Triplet energy transfer (TT), a key process in molecular and organic electronics, generally occurs by either strongly distance-dependent single-step tunneling or weakly distance-dependent multistep hopping. We have synthesized a series of π-stacked molecules consisting of a benzophenone donor, one to three fluorene bridges, and a naphthalene acceptor, and studied the rate of TT from benzophenone to naphthalene across the fluorene bridge using femtosecond transient absorption spectroscopy. We show that the dominant TT mechanism switches from tunneling to wire-like hopping between bridge lengths 1 and 2. The crossover observed for TT can be determined by direct observation of the bridge-occupied state.

A “Radical Dance” in Thiamin Biosynthesis: Mechanistic Analysis of the Bacterial Hydroxymethylpyrimidine Phosphate Synthase

Thiamin pyrophosphate is an important cofactor in all forms of life, where it plays a central role in the stabilization of the acyl-carbanion biosynthon1, 2. Its biosynthesis involves separate synthesis of the thiazole and the pyrimidine hererocycles, which are then linked to form the cofactor. Thiamin-thiazole biosynthesis is relatively well-understood3-7. In prokaryotes, 1-deoxy-D-xylulose-5-phosphate, cysteine and glycine or tyrosine are utilized by five proteins to construct the thiazole moiety, whereas in Saccharomyces cerevisiae, just one gene product converts NAD and glycine to thiazole, obtaining sulfur from a source yet unknown. In comparison, the mechanistic understanding of thiamin-pyrimidine (HMP) biosynthesis, in both prokaryotes and eukaryotes, is still at an early stage. In yeast, a single gene product THI5p is implicated in HMP biosynthesis from PLP and histidine, however this reaction has not yet been successfully reconstituted in vitro. In bacteria and plants HMP-P synthase (ThiC) catalyzes the conversion of aminoimidazole ribonucleotide (AIR 1), an intermediate in the purine nucleotide biosynthesis pathway, to hydroxylmethyl pyrimidine phosphate (HMP-P 2)8. In vivo and in vitro studies on the reaction catalyzed by ThiC, using labeled AIR, have revealed the involvement of a rearrangement reaction of remarkable complexity (Figure 1A)9. The ThiC catalyzed reaction has recently been reconstituted in a defined biochemical system. Spectroscopic, structural and biochemical studies established this enzyme as a unique member of the [4Fe-4S] cluster dependent radical-SAM superfamily10-11.

Radical S-Adenosylmethionine (SAM) Enzymes in Cofactor Biosynthesis: A Treasure Trove of Complex Organic Radical Rearrangement Reactions

In this minireview, we describe the radical S-adenosylmethionine enzymes involved in the biosynthesis of thiamin, menaquinone, molybdopterin, coenzyme F420, and heme. Our focus is on the remarkably complex organic rearrangements involved, many of which have no precedent in organic or biological chemistry.

Electrochemistry and Electrogenerated Chemiluminescence of π‑Stacked Poly(fluorenemethylene) Oligomers. Multiple, Interacting Electron Transfers

The electrochemistry, spectroscopy, and electrogenerated chemiluminescence (ECL) of a series of π-stacked poly(fluorenemethylene) oligomers (Fn, n = 1-6) were investigated. The pendant cofacially oriented fluorene moieties are essentially in contact with each other by Van der Waals interaction promoting electronic delocalization in these species. All six compounds give successive cyclic voltammetric one-electron (1e) oxidations in 1:1 acetonitrile/benzene (MeCN/Bz), and the multiple 1e transfer properties of all these compounds were confirmed by chronoamperometric experiments with an ultramicroelectrode and digital simulations. The potentials for oxidation of the successive 1e transfers can be explained in terms of electrostatic interactions among the fluorenes. The monomer (F1) shows one irreversible wave, while F2 shows two reversible 1e waves. F3 shows only two reversible 1e oxidation waves, which is consistent with the large energy to remove a third electron because of the greater electrostatic repulsion, so the third wave is shifted toward more positive potentials. Both F4 and F5 show three reversible 1e oxidation waves, while F6 shows four reversible 1e waves. The removal of the first electron from an oligomer becomes easier as n increases. The stability of the radical cations also increases with n. The removal of consecutive electrons from Fn can be correlated with the distance between fluorene moieties. No reduction peaks were observed except for some broad ones at ~-3.2 V vs SCE in THF, which is consitent with the wide highest occupied molecular orbital-lowest unoccupied molecular orbital gap in these compounds (absorbance at about 300 nm). No characteristic annihilation ECL signal was observed for these compounds in 1:1 MeCN/Bz mixed solvent. However, the ECL of F6 in the presence of the coreactant C(2)O(4)(2-) showed a long-wavelength ECL emission that was proposed to be electrolyzed byproduct from the radical cation.

Non-canonical active site architecture of the radical SAM thiamin pyrimidine synthase

Radical S-adenosylmethionine (SAM) enzymes use a [4Fe-4S] cluster to generate a 5′-deoxyadenosyl radical. Canonical radical SAM enzymes are characterized by a β-barrel-like fold and SAM anchors to the differentiated iron of the cluster, which is located near the amino terminus and within the β-barrel, through its amino and carboxylate groups. Here we show that ThiC, the thiamin pyrimidine synthase in plants and bacteria, contains a tethered cluster-binding domain at its carboxy terminus that moves in and out of the active site during catalysis. In contrast to canonical radical SAM enzymes, we predict that SAM anchors to an additional active site metal through its amino and carboxylate groups. Superimposition of the catalytic domains of ThiC and glutamate mutase shows that these two enzymes share similar active site architectures, thus providing strong evidence for an evolutionary link between the radical SAM and adenosylcobalamin-dependent enzyme superfamilies.

Catalysis of a Flavoenzyme-Mediated Amide Hydrolysis

A new pyrimidine catabolic pathway (the Rut pathway) was recently discovered in Escherichia coli K12. In this pathway, uracil is converted to 3-hydroxypropionate, ammonia, and carbon dioxide. The seven-gene Rut operon is required for this conversion. Here we demonstrate that the flavoenzyme RutA catalyzes the initial uracil ring-opening reaction to give 3-ureidoacrylate. This reaction, while formally a hydrolysis reaction, proceeds by an oxidative mechanism initiated by the addition of a flavin hydroperoxide to the C4 carbonyl. While peroxide-catalyzed amide hydrolysis has chemical precedent, we are not aware of a prior example of analogous chemistry catalyzed by flavin hydroperoxides. This study further illustrates the extraordinary catalytic versatility of the flavin cofactor.

In vitro reconstitution of the radical S-adenosylmethionine enzyme MqnC involved in the biosynthesis of futalosine-derived menaquinone.

The radical S-adenosylmethionine enzyme MqnC catalyzes conversion of dehypoxanthine futalosine (DHFL) to the unique spiro compound cyclic DHFL in the futalosine pathway for menaquinone biosynthesis. This study describes the in vitro reconstitution of [4Fe-4S] cluster-dependent MqnC activity and identifies the site of abstraction of a hydrogen atom from DHFL by the adenosyl radical.

Identification of the product of Toxoflavin lyase: degradation via a Baeyer-Villiger oxidation

Toxoflavin (an azapteridine) is degraded to a single product by toxoflavin lyase (TflA) in a reaction dependent on reductant, Mn(II), and oxygen. The isolated product was fully characterized by NMR and MS and was identified as a triazine in which the pyrimidine ring was oxidatively degraded. A mechanism for toxoflavin degradation based on the identification of the enzymatic product and the recently determined crystal structure of toxoflavin lyase is proposed.

Anaerobic 5-Hydroxybenzimidazole Formation from Aminoimidazole Ribotide: An Unanticipated Intersection of Thiamin and Vitamin B12 Biosynthesis

Comparative genomics of the bacterial thiamin pyrimidine synthase (thiC) revealed a paralogue of thiC (bzaF) clustered with anaerobic vitamin B12 biosynthetic genes. Here we demonstrate that BzaF is a radical S-adenosylmethionine enzyme that catalyzes the remarkable conversion of aminoimidazole ribotide (AIR) to 5-hydroxybenzimidazole (5-HBI). We identify the origin of key product atoms and propose a reaction mechanism. These studies represent the first step in solving a long-standing problem in anaerobic vitamin B12 assembly and reveal an unanticipated intersection of thiamin and vitamin B12 biosynthesis.

A Remarkable Oxidative Cascade That Replaces the Riboflavin C8 Methyl with an Amino Group during Roseoflavin Biosynthesis

Roseoflavin is a naturally occurring riboflavin analogue with antibiotic properties. It is biosynthesized from riboflavin in a reaction involving replacement of the C8 methyl with a dimethylamino group. Herein we report the identification of a flavin-dependent enzyme that converts flavin mononucleotide (FMN) and glutamate to 8-amino-FMN via the intermediacy of 8-formyl-FMN. A mechanistic proposal for this remarkable transformation is proposed.