Evidence for the Prerequisite Formation of Phenoxyl Radicals in Radical-Mediated Peptide Tyrosine Nitration In Vacuo

The elementary mechanism of radical-mediated peptide tyrosine nitration, which is a hallmark of post-translational modification of proteins under nitrative stress in vivo, has been elucidated in detail by using an integrated approach that combines the gas-phase synthesis of prototypical molecular tyrosine-containing peptide radical cations, ion–molecule reactions, and isotopic labeling experiments with DFT calculations. This reaction first involves the radical recombination of ^.NO₂ towards the prerequisite phenoxyl radical tautomer of a tyrosine residue, followed by proton rearrangements, finally yielding the stable and regioselective 3-nitrotyrosyl residue product. In contrast, nitration with the π-phenolic radical cation tautomer is inefficient. This first direct experimental evidence for the elementary steps of the radical-mediated tyrosine nitration mechanism in the gas phase provides a fundamental insight into the regioselectivity of biological tyrosine ortho-nitration.     

Synthesis of High Molecular Weight Polyesters via In Vacuo Dehydrogenation Polymerization of Diols

The Milstein catalyst has proven to be highly effective for the conversion of alcohols to esters, as well as alcohols and amines to amides and polyamides. We have recently found that the catalyst's range can be extended to very efficient in vacuo dehydrogenation polymerization of α,ω‐diols to generate polyesters. The gaseous hydrogen byproduct that is produced is easily removed to drive the equilibrium toward product, which leads to the formation of high molecular weight polymer ( up to 145 000 g mol⁻¹). This optimized methodology works well to polymerize diols with a spacer of six carbons or more. Diols with fewer carbons are cyclized to lactone; the dividing point is the dehydrogenation of 1,5‐pentanediol, which leads to a mixture of polyester and lactone. Reported herein is the synthesis and characterization of five aliphatic polyesters prepared via this novel dehydrogenation polymerization approach.     

Persistent Room-Temperature Radicals from Anionic Naphthalimides: Spin Pairing and Supramolecular Chemistry

N-Substituted naphthalimides (NNIs) have been shown to exhibit highly efficient and persistent room-temperature phosphorescence from an NNI-localized triplet excited state, when the N-substitution is a sufficiently strong donor and mediates an intramolecular charge-transfer (ICT) state upon photo-excitation. This work shows that, when the electron-donating ability of the N-substitution is further increased in the presence of a carbanion or phenoxide, spontaneous electron transfer (ET) occurs and results in radical anions, verified with electron-paramagnetic resonance (EPR) spectroscopy. However, the EPR-active anion is surprisingly persistent and impervious to nucleophilic and radical reactions under anionic conditions. The stability is thought to originate from an intramolecular spin pairing between the N-donor and the NI acceptor post ET, which is demonstrated in supramolecular chemistry.     

Ground and Excited-Electronic-State Dissociations of Hydrogen-Rich and Hydrogen-Deficient Tyrosine Peptide Cation Radicals

We report a comprehensive study of collision-induced dissociation (CID) and near-UV photodissociation (UVPD) of a series of tyrosine-containing peptide cation radicals of the hydrogen-rich and hydrogen-deficient types. Stable, long-lived, hydrogen-rich peptide cation radicals, such as [AAAYR + 2H]^+● and several of its sequence and homology variants, were generated by electron transfer dissociation (ETD) of peptide-crown-ether complexes, and their CID-MS³ dissociations were found to be dramatically different from those upon ETD of the respective peptide dications. All of the hydrogen-rich peptide cation radicals contained major (77%–94%) fractions of species having radical chromophores created by ETD that underwent photodissociation at 355 nm. Analysis of the CID and UVPD spectra pointed to arginine guanidinium radicals as the major components of the hydrogen-rich peptide cation radical population. Hydrogen-deficient peptide cation radicals were generated by intramolecular electron transfer in Cu^II(2,2′:6′,2″-terpyridine) complexes and shown to contain chromophores absorbing at 355 nm and undergoing photodissociation. The CID and UVPD spectra showed major differences in fragmentation for [AAAYR]^+● that diminished as the Tyr residue was moved along the peptide chain. UVPD was found to be superior to CID in localizing C_α-radical positions in peptide cation radical intermediates.

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Graphical Abstract ?

     

Basic Principles of Protease-Catalyzed Peptide Bond Formation

The stereospecific formation of peptide bonds under mild conditions and without side reactions is still a formidable task in peptide synthesis. One approach that springs to mind, namely the use of the naturally occurring catalyst involved in the biosynthesis of proteins, the ribosomal peptidyl transferase, cannot be realized in practice. The fact, however, that the natural cleavage of proteins is carried out by other enzymes, namely the proteases, together with the reversibility of these cleavage reactions in principle, has led to an interesting synthetic concept. Proteases normally catalyze the enzymatic degradation of proteins and peptides by hydrolytic cleavage of the peptide bond in an exergonic reaction. The use of physicochemical principles in order to influence the equilibrium, the concentration of products, and the kinetic parameters of the reaction results in the successful application of the catalytic properties of proteases to peptide synthesis. The purpose of this review is to describe and summarize the methods used in such approaches and to attempt a systematic categorization. The principles are applied to the synthesis of such practically relevant products as aspartame and human insulin.     

Formation of Mixed Ionic Complementary Peptide Fibrils

Summary: Two peptides (FEFEFKFK and HHHHHHFEFEFKFK) have been synthesised and their phase diagrams mapped out as a function of concentration and temperature. Both peptides formed self-supporting fibrillar hydrogels above a similar critical molar gelation concentration with a fibril diameter corresponding to the length of the fully stretched monomer. Mixing the peptides in a 9:1 ratio of FEFEFKFK to histadine functionalized FEFEFKFK in the presence of nanogold particles (binds to the histadine groups) resulted in thicker fibres suggesting that two fibrils associate together to form a fibre. TEM studies revealed that the gold particles were distributed throughout the hydrogel and adjacent to both sides of the fibrillar structures with an average distance between particles of 21 nm. It is postulated that the peptides form anti-parallel beta sheet fibrils that associate together via π-stacking interactions between the imidazole side chains of the histadine groups to form a fibre, where on average 1 in every 44 peptides is functionalized.     

Formation of Diverse Mesophases Templated by a Diprotic Anionic Surfactant

The synthesis system for mesophase formation, using the diprotic anionic surfactant N‐myristoyl‐L ‐glutamic acid (C₁₄GluA) as the structure‐directing agent (SDA) and N‐trimethoxylsilylpropyl‐N,N,N‐trimethylammonium chloride (TMAPS) as the co‐structure‐directing agent (CSDA), has been investigated and a full‐scaled synthesis‐field diagram is presented. In this system we have obtained mesophases including three‐dimensional (3D) micellar cubic Fm m, Pm n, Fd m, micellar tetragonal P4₂/mnm, two‐dimensional (2D) hexagonal p6mm and bicontinuous cubic Pn m, by varying the C₁₄GluA/NaOH/TMAPS composition ratios. From the diagram it can be concluded that the mesophase formation is affected to a high degree by the organic/inorganic‐interface curvature and the mesocage–mesocage electrostatic interaction. Bicontinuous cubic and 2D‐hexagonal phases were found in the low organic/inorganic‐interface curvature zones, whereas micellar cubic and tetragonal mesophases were found in the high organic/inorganic‐interface curvature zones. Formation of cubic Fm m and tetragonal P4₂/mnm was favoured in highly alkaline zones with strong mesocage–mesocage interactions, and formation of cubic Pm n and Fd m was favoured with moderate mesocage–mesocage interactions in the less alkaline zones of the diagram.     

Phosphorus Compounds for the Formation of Unprecedented Anionic Zirconium Complexes

Using the nucleophilic ability of tertiary phosphines we have been able to prepare a large variety of unprecedented stable phosphonium anionic zirconocene(IV) complexes.     

Formation of CC Bonds by Addition of Free Radicals to Alkenes

There are many reactions in which CC bonds are formed by addition of free radicals to alkenes. Information about the mechanism is important for the synthesis of specific target molecules. The rate of addition of alkyl radicals to alkenes is controlled by steric and polar effects. The stabilities of the educts and products are of only limited importance, since the transition states for these exothermic reactions occur very early on the reaction coordinate. Variations in reactivity and selectivity can be described using frontier orbital theory: for nucleophilic radicals the dominant interactions are those between SOMO's and LUMO's, and for electrophilic radicals those between SOMO's and HOMO's. The large differences in the steric effects of α - and β- substituents of alkenes can be explained by postulating an unsymmetrical transition state— the radical approaches one of the C atoms preferentially. Regioand stereoselectivities can be predicted and are determined, in general, by steric effects.     

Glycine Peptide Bond Formation Catalyzed by Faujasite

The catalysis of peptide bond formation between two glycine molecules on H‐FAU zeolite was computationally studied by the M08‐HX density functional. Two reaction pathways, the concerted and the stepwise mechanism, starting from three differently adsorbed reactants, amino‐bound, carboxyl‐bound, and hydroxyl‐bound, are studied. Adsorption energies, activation energies, and reaction energies, as well as the corresponding intrinsic rate constants were calculated. A comparison of the computed energetics of the various reaction paths for glycine indicates that the catalyzed reaction proceeds preferentially via the concerted reaction mechanism of the hydroxyl‐bound configuration. This involves an eight‐membered ring of the transition structure instead of the four‐membered ring of the others. The step from the amino‐bound configuration to glycylglycine is the rate‐determining step of the concerted mechanism. It has an estimated activation energy of 51.2 kcal mol^?1. Although the catalytic reaction can also occur via the stepwise reaction mechanism, this path is not favored.     

Formation and Dissociation of Phosphorylated Peptide Radical Cations

In this study, we generated phosphoserine- and phosphothreonine-containing peptide radical cations through low-energy collision-induced dissociation (CID) of the ternary metal?Cligand phosphorylated peptide complexes [Cu^II(terpy) _p M]^·2+ and [Co^III(salen) _p M]^·+ [ _p M: phosphorylated angiotensin III derivative; terpy: 2,2':6',2''-terpyridine; salen: N,N '-ethylenebis(salicylideneiminato)]. Subsequent CID of the phosphorylated peptide radical cations ( _p M^·+) revealed fascinating gas-phase radical chemistry, yielding (1) charge-directed b- and y-type product ions, (2) radical-driven product ions through cleavages of peptide backbones and side chains, and (3) different degrees of formation of [M ?C H₃PO₄]^·+ species through phosphate ester bond cleavage. The CID spectra of the _p M^·+ species and their non-phosphorylated analogues featured fragment ions of similar sequence, suggesting that the phosphoryl group did not play a significant role in the fragmentation of the peptide backbone or side chain. The extent of neutral H₃PO₄ loss was influenced by the peptide sequence and the initial sites of the charge and radical. A preliminary density functional theory study, at the B3LYP 6-311++G(d,p) level of theory, of the neutral loss of H₃PO₄ from a prototypical model??N-acetylphosphorylserine methylamide??revealed several factors governing the elimination of neutral phosphoryl groups through charge- and radical-induced mechanisms.     

Calcium Hydride Reactivity: Formation of an Anionic N-Heterocyclic Olefin Ligand

An anionic N-heterocyclic olefin ligand was serendipitously obtained by reaction of an amidinate calcium hydride complex with 1,3-dimethyl-2-methyleneimidazole (NHO). Instead of anticipated addition to the polarized C=CH₂ bond to form an unstabilized alkylcalcium complex, deprotonation of the NHO ligand in the backbone was observed. Preference for deprotonation versus addition is explained by loss of aromaticity in the latter conversion. Theoretical calculations demonstrate the substantially increased ylidic character of this anionic NHO ligand which, like N-heterocyclic dicarbenes, shows strong bifunctional coordination.     

Hydroxyl Radicals Formation in Dielectric Barrier Discharge During Decomposition of Toluene

A method based on high performance liquid chromatography (HPLC), has been developed to measure hydroxyl radical (^·OH) in plasma reactors. The determination was performed indirectly by detecting the products of the reaction of ^·OH with salicylic acid (SAL). The applicability, and effect of time, specific input energy (SIE), relative humidity (RH), catalyst were investigated. It was found that 3 h was the optimal trapping time; concentration of ^·OH was (5.9–23.6) × 10¹³ radicals/cm³ at SIE range. The highest ^·OH yield and toluene removal efficiency (η) were achieved with a RH of 20%. With MnO _x , η was two times that without catalyst, while ^·OH yield in gas stream was one-sixth that without catalyst. However, if summed with ^·OH adsorbed on catalyst surface, the total ^·OH yield was the same as without catalyst. Experiments performed with/without toluene allowed to determine the role of ^·OH on decomposition of toluene in air plasma.     

Complexation of Nitroxide Radicals with Cyclodextrins: Kinetics of Crystal Complex Formation

Using the ESR method in combination with the stop-flow technique the kinetics of crystal complex formation of hydrophobic nitroxide radicals with cyclodextrins has been studied upon mixing cyclodextrin aqueous solutions with nitroxides emulsified in aqueous milieu. The - and -cyclodextrins and the piperidine nitroxide radicals with a OC(O)C_mH_2m+i (m = 7, 10, and 17) substituent in a para-position were used. It was established that the complexation process in the system emulsion of probe–cyclodextrin solution consists of two mainstages. The first stage is the transfer of probes from drops into aqueous solution and the formation of complexes RC_n, where R is the probe, C is cyclodextrin, n = 1, 2 and 3. The second stage is crystal complex formation and growth from solution of RC_n complexes. The results obtained indicate that mainly RC₃ complexes take part in crystallization. It was observed that the characteristic time of crystallization is approximately inversely proportional to the concentration of RC₃ complexes.Equilibrium constants of the processes R + C RC, RC + C RC₂ have been determined. It was found that complexation and further crystallization lead to the formation of monodispersed microcrystals.     

Selective Formation of Indene through the Reaction of Benzyl Radicals with Acetylene

The combustion of fossil fuels forms polycyclic aromatic hydrocarbons (PAHs) composed of five‐ and six‐ membered aromatic rings, such as indene (C₉H₈), which are carcinogenic, mutagenic, and deleterious to the environment. Indene, the simplest PAH with single five‐ and six‐membered rings, has been predicted theoretically to be formed through the reaction of benzyl radicals with acetylene. Benzyl radicals are found in significant concentrations in combustion flames, owing to their highly stable aromatic and resonantly stabilized free‐radical character. We provide compelling experimental evidence that indene is synthesized through the reaction of the benzyl radical (C₇H₇) with acetylene (C₂H₂) under combustion‐like conditions at 600 K. The mechanism involves an initial addition step followed by cyclization and aromatization through atomic hydrogen loss. This reaction was found to form the indene isomer exclusively, which, in conjunction with the high concentrations of benzyl and acetylene in combustion environments, indicates that this pathway is the predominant route to synthesize the prototypical five‐ and six‐membered PAH.     

正负离子表面活性剂混合体系中高稳定性囊泡的形成

对总浓度为0.01 mol/L，摩尔比为2：1的十二烷基硫酸钠/溴化十二烷基三乙 铵的正负离子表面活性剂混合体系形成的囊泡的稳定性进行了研究。发现这一体系 形成的囊泡在长放置（5个月）后依然存在。在加入较大量的无机盐（0.15 mol/L NaBr）、较大幅度pH变化（pH = 2～12）、温度变化（从80 ℃到-22 ℃）情况下 体系中的囊泡依然呈现出优异的稳定性。在非水溶剂乙醇（100%）中这类正负离子 表面活性剂仍然可以形成囊泡。