A Non-canonical PDK1-RSK Signal Diminishes Pro-caspase-8-Mediated Necroptosis Blockade.

Necroptosis induction in vitro often requires caspase-8 (Casp8) inhibition by zVAD because pro-Casp8 cleaves RIP1 to disintegrate the necrosome. It has been unclear how the Casp8 blockade of necroptosis is eliminated naturally. Here, we show that pro-Casp8 within the necrosome can be inactivated by phosphorylation at Thr265 (pC8T265). pC8T265 occurs in vitro in various necroptotic cells and in the cecum of TNF-treated mice. p90 RSK is the kinase of pro-Casp8. It is activated by a mechanism that does not need ERK but PDK1, which is recruited to the RIP1-RIP3-MLKL-containing necrosome. Phosphorylation of pro-Casp8 at Thr265 can substitute for zVAD to permit necroptosis in vitro. pC8T265 mimic T265E knockin mice are embryonic lethal due to unconstrained necroptosis, and the pharmaceutical inhibition of RSK-mediated pC8T265 diminishes TNF-induced cecum damage and lethality in mice by halting necroptosis. Thus, phosphorylation of pro-Casp8 at Thr265 by RSK is an intrinsic mechanism for passing the Casp8 checkpoint of necroptosis.

Targeting the PHF8/YY1 axis suppresses cancer cell growth through modulation of ROS.

High levels of mitochondrial reactive oxygen species (mROS) are linked to cancer development, which is tightly controlled by the electron transport chain (ETC). However, the epigenetic mechanisms governing ETC gene transcription to drive mROS production and cancer cell growth remain to be fully characterized. Here, we report that protein demethylase PHF8 is overexpressed in many types of cancers, including colon and lung cancer, and is negatively correlated with ETC gene expression. While it is well known to demethylate histones to activate transcription, PHF8 demethylates transcription factor YY1, functioning as a co-repressor for a large set of nuclear-coded ETC genes to drive mROS production and cancer development. In addition to genetically ablating PHF8, pharmacologically targeting PHF8 with a specific chemical inhibitor, iPHF8, is potent in regulating YY1 methylation, ETC gene transcription, mROS production, and cell growth in colon and lung cancer cells. iPHF8 exhibits potency and safety in suppressing tumor growth in cell-line- and patient-derived xenografts in vivo. Our data uncover a key epigenetic mechanism underlying ETC gene transcriptional regulation, demonstrating that targeting the PHF8/YY1 axis has great potential to treat cancers.

Carbon-Atom Exchange between [MC2]+ (M = Os and Ir) and Methane: on the Thermodynamic and Dynamic Aspects.

Gas-phase reactions of [OsC2]+ and [IrC2]+ with methane at ambient temperature have been studied using quadrupole-ion trap mass spectrometry combined with quantum chemical calculations. Both [OsC2]+ and [IrC2]+ undergo carbon-atom exchange reactions with methane. The associated mechanisms for the two systems are found to be similar. The differences in the rates of carbon isotope exchange reactions of methane with [MC2]+ (M = Os and Ir) are explained by several factors like the energy barrier for the initial H3C-H bond breaking processes, the molecular dynamics, orbital interactions, and the H-binding energies of the pivotal steps. Besides, the number of participating valence orbitals might be one of the keys to regulate the rate in the key step. The present findings may provide useful ideas and inspiration for designing similar processes.

Nonlinear differential equations and their application to evaluating the integrated impacts of multiple parameters on the biochemical safety of drinking water.

The present study aimed to narrow such gaps by applying nonlinear differential equations to biostability in drinking water. Biostability results from the integrated dynamics of nutrients and disinfectants. The linear dynamics of biostability have been well studied, while there remain knowledge gaps concerning nonlinear effects. The nonlinear effects are explained by phase plots for specific scenarios in a drinking water system, including continuous nutrient release, flush exchange with the adjacent environment, periodic pulse disinfection, and periodic biofilm development. The main conclusions are, (1) The correlations between the microbial community and nutrients go through phases of linear, nonlinear, and chaotic dynamics. Disinfection breaks the chaotic phase and returns the system to the linear phase, increasing the microbial growth potential. (2) Post-disinfection after multiple microbial peaks produced via metabolism can increase disinfection efficiency and decrease the risks associated with disinfectant byproduct risks. This can provide guidelines for optimizing the disinfection strategy, according to the long-term water safety target or a short management. Limited disinfection and ultimate disinfection may be more effective and have low chemical risk, facing longer stagnant conditions. (3) Periodic biofilm formation and biofilm detachment increase the possibility of uncertainty in the chaotic phase. For future study, nonlinear differential equation models can accordingly be applied at the molecular and ecological levels to further explore more nonlinear regulation mechanisms.

On the performance of the M-C (M = Fe, Ru, Os) unit toward methane activation.

Gas-phase reactions of [MC]+ (M = Os and Ru) with methane at ambient temperature have been studied by using quadrupole-ion trap (Q-IT) mass spectrometry combined with quantum chemical calculations. Theoretical calculations reveal the influence of electronic signatures and that it is the energy gap of the associated frontier molecular orbitals that dominates the ability of the cluster in the initial H3C-H bond breaking. By extension, a theoretical consideration upon changing the ligand from carbide to carbyne and eventually to carbene reveals that the reactivities of the M-complex (M = Os, Ru and Fe) are determined by the energy gap of the involved orbitals. In addition, a few factors like the dipole moment, spin density and charge distributions influence the orbital energy gap to different extents. Thus, altering the local structure of the active center to modulate the orbital distribution may be a possible means of regulation of the activity.

Iridium Dimer Anion-Mediated C≡C Triple Bond Cleavage and Successive Dehydrogenation of Acetylene in the Gas Phase.

The reactions of the iridium dimer anion [Ir2]- with acetylene have been studied by mass spectrometry in the gas phase, which indicate that the [Ir2]- anion can consecutively react with C2H2 molecules to form the [Ir2C2x]- (x = 1, 2) and [Ir2C2yH2]- (y = 3-5) anions as major products with the successive release of H2 molecules at room temperature. The reactions are confirmed by the reactions of the mass-selected product [Ir2C2]- anion with C2H2 to produce [Ir2C4]- and [Ir2C2yH2]- (y = 3-5). Photoelectron spectra and quantum chemistry calculations confirm that the [Ir2C2x]- (x = 1, 2) product anions possess cyclic [Ir(µ-C)2Ir]- and [Ir(µ-C)(µ-C3)Ir]- structures, implying that the robust C≡C triple bond of acetylene can be completely cleaved by the [Ir2]- anion.

Reactions of Transition-Metal Carbyne Cations with Ethylene in the Gas Phase.

The reactions of iridium- and osmium-carbyne hydride cations [HIrCH]+ and [HOsCH]+ with ethylene have been studied using mass spectrometry with isotopic-labeling in the gas phase. The carbyne reactivity is compared with that of the rhodium, cobalt, and iron analogues [TMCH2]+ (TM = Fe, Co, and Rh), which were determined to have the carbene structures. Besides the cycloaddition/dehydrogenation reaction in forming the [TMC3H4]+ + H2 (TM = Ir and Os) products, a second reaction pathway producing the [TMC2H2]+ ion and CH4 via triple hydrogen atom transfer reactions to the carbyne carbon is observed to be the major channel. The latter channel is not observed in the rhodium, cobalt, and iron carbene cation reactions. Quantum-chemical calculations indicate that the distinct reactivity is not due to different initial structures of the reactants. Both reaction channels are predicted to be thermodynamically exothermic and kinetically facile for the carbyne cations, and the reactions proceed with the initial formation of a carbene intermediate via hydride-carbyne coupling. The latter channel is also exothermic but kinetically unfavorable for the rhodium, cobalt, and iron carbene cations.

Arginine methylation of HSP70 regulates retinoid acid-mediated RARß2 gene activation.

Although "histone" methyltransferases and demethylases are well established to regulate transcriptional programs and to use nonhistone proteins as substrates, their possible roles in regulation of heat-shock proteins in the nucleus have not been investigated. Here, we report that a highly conserved arginine residue, R469, in HSP70 (heat-shock protein of 70 kDa) proteins, an evolutionarily conserved protein family of ATP-dependent molecular chaperone, was monomethylated (me1), at least partially, by coactivator-associated arginine methyltransferase 1/protein arginine methyltransferase 4 (CARM1/PRMT4) and demethylated by jumonji-domain-containing 6 (JMJD6), both in vitro and in cultured cells. Functional studies revealed that HSP70 could directly regulate retinoid acid (RA)-induced retinoid acid receptor ß2 (RARß2) gene transcription through its binding to chromatin, with R469me1 being essential in this process. HSP70's function in gene transcriptional regulation appears to be distinct from its protein chaperon activity. R469me1 was shown to mediate the interaction between HSP70 and TFIIH, which involves in RNA polymerase II phosphorylation and thus transcriptional initiation. Our findings expand the repertoire of nonhistone substrates targeted by PRMT4 and JMJD6, and reveal a new function of HSP70 proteins in gene transcription at the chromatin level aside from its classic role in protein folding and quality control.

Striking Doping Effects on Thermal Methane Activation Mediated by the Heteronuclear Metal Oxides [XAlO4 ].+ (X=V, Nb, and Ta).

The thermal reactivity of the heteronuclear metal-oxide cluster cations [XAlO4 ].+ (X=V, Nb, and Ta) towards methane has been studied by using mass spectrometry in conjunction with quantum mechanical calculations. Experimentally, a hydrogen-atom transfer (HAT) from methane is mediated by all the three oxide clusters at ambient conditions. However, [VAlO4 ].+ is unique in that this cluster directly transforms methane into formaldehyde. The absence of this reaction for the Nb and Ta analogues demonstrates a striking doping effect on the chemoselectivity in the conversion of methane. Mechanistic aspects of the two reactions have been elucidated by quantum-chemical calculations. The HAT reactivity can be attributed to the significant spin density localized at the terminal oxygen atom (Ot.- ) of the cluster ions, while the ionic/covalent character of the Lewis acid-base unit [X-Ob ] plays a crucial role for the generation of formaldehyde. The mechanistic insight derived from this combined experimental/computational investigation may provide guidance for a more rational design of catalysts.

Unravelling Mechanistic Aspects of the Gas-Phase Ethanol Conversion: An Experimental and Computational Study on the Thermal Reactions of MO2 (+) (M=Mo, W) with Ethanol.

The ion/molecule reactions of molybdenum and tungsten dioxide cations with ethanol have been studied by Fourier transform ion-cyclotron resonance mass spectrometry (FT-ICR MS) and density functional theory (DFT) calculations. Dehydration of ethanol has been found as the dominant reaction channel, while generation of the ethyl cation corresponds to a minor product. Cleary, the reactions are mainly governed by the Lewis acidity of the metal center. Computational results, together with isotopic labeling experiments, show that the dehydration of ethanol can proceed either through a conventional concerted [1,2]-elimination mechanism or a step-wise process; the latter occurs via a hydroxyethoxy intermediate. Formation of C2 H5 (+) takes place by transfer of OH(-) from ethanol to the metal center of MO2 (+) . The molybdenum and tungsten dioxide cations exhibit comparable reactivities toward ethanol, and this is reflected in similar reaction rate constants and branching ratios.

Efficient Room-Temperature, Au(+)-Mediated Coupling of a Carbene Ligand with Methane To Generate C2Hx (x = 4, 6).

The thermal reactions of the closed-shell, "naked" gold-carbene complex [Au(CH2)](+) with methane have been explored by using FTICR mass spectrometry complemented by quantum chemical (QC) calculations at the CCSD(T)//BMK level of theory. Mechanistic aspects for this unprecedentedly efficient carbene insertion in the C-H bond of methane have been addressed and the origin of the counterintuitive high reactivity of [Au(CH2)](+) towards this most inert hydrocarbon is discussed.

A Tin Analogue of Carbenoid: Isolation and Reactivity of a Lithium Bis(imidazolin-2-imino)stannylenoid.

The lithium bis(imino)stannylenoid (NIPr)2 Sn(Li)Cl (1; NIPr=bis(2,6-diisopropylphenyl)imidazolin-2-imino) was prepared by the reaction of LiNIPr with 0.5 equiv of SnCl2 ⋅diox (diox=1,4-dioxane) and the ambiphilic character of the compound was demonstrated by investigations into its reactivity. Treatment of 1 with I2 or MeI yielded the oxidative addition products (NIPr)2 SnI2 and (NIPr)2 Sn(Me)I, respectively. In contrast, the reaction of 1 with one equivalent of Me3 SiCl resulted in the formation of Me3 SiNIPr and the chlorostannylene dimer [NIPrSnCl]2 . Moreover, the substitution reaction of compound 1 with MeLi led to the formation of the methyl-substituted stannate (NIPr)2 Sn(Li)Me.

"Stripping" the carbon atom of methyl halide by a cationic holmium complex: a gas-phase study.

Mechanistic aspects of an unusual reaction of [HoC6 H4 S](+) with CH3 X (X=Cl, Br, I) have been investigated using Fourier-transform ion cyclotron resonance mass spectrometry combined with density functional theory (DFT) calculations. In this thermal process, all four bonds of the methyl halides are cleaved.

Carbon-Atom Extrusion from Halobenzenes and Its Coupling with a Methylene Ligand to Form Acetylene.

Mechanistic aspects of an unusual gas-phase reaction of [LaCH2](+) with halobenzenes have been investigated using Fourier-transform ion cyclotron resonance (FTICR) mass spectrometry combined with density functional theory (DFT) calculations. In this thermal process a carbon-atom from the benzene ring, most likely the ipso-position, and the carbene ligand are coupled to form C2H2.

High reactivity of nanosized niobium oxide cluster cations in methane activation: A comparison with vanadium oxides.

The reactions between methane and niobium oxide cluster cations were studied and compared to those employing vanadium oxides. Hydrogen atom abstraction (HAA) reactions were identified over stoichiometric (Nb2O5)N(+) clusters for N as large as 14 with a time-of-flight mass spectrometer. The reactivity of (Nb2O5)N(+) clusters decreases as the N increases, and it is higher than that of (V 2O5)N(+) for N ≥ 4. Theoretical studies were conducted on (Nb2O5)N(+) (N = 2-6) by density functional calculations. HAA reactions on these clusters are all favorable thermodynamically and kinetically. The difference of the reactivity with respect to the cluster size and metal type (Nb vs V) was attributed to thermodynamics, kinetics, the electron capture ability, and the distribution of the unpaired spin density. Nanosized Nb oxide clusters show higher HAA reactivity than V oxides, indicating that niobia may serve as promising catalysts for practical methane conversion.

On the Mechanisms of Hydrogen-Atom Transfer from Water to the Heteronuclear Oxide Cluster [Ga2 Mg2 O5 ](.+) : Remarkable Electronic Structure Effects.

Mechanistic insight into the homolytic cleavage of the O-H bond of water by the heteronuclear oxide cluster [Ga2 Mg2 O5 ](.+) has been derived from state-of-the-art gas-phase experiments in conjunction with quantum chemical calculations. Three pathways have been identified computationally. In addition to the conventional hydrogen-atom transfer (HAT) to the radical center of a bridging oxygen atom, two mechanistically distinct proton-coupled electron-transfer (PCET) processes have been identified. The energetically most favored path involves initial coordination of the incoming water ligand to a magnesium atom followed by an intramolecular proton transfer to the lone-pair of the bridging oxygen atom. This step, which is accomplished by an electronic reorganization, generates two structurally equivalent OH groups either of which can be liberated, in agreement with labeling experiments.

Distinct mechanistic differences in the hydrogen-atom transfer from methane and water by the heteronuclear oxide cluster [Ga2 MgO4](.).

The thermal reactions of the heteronuclear oxide cluster [Ga2 MgO4 ](.+) with methane and water have been studied using state-of-the-art gas-phase experiments in conjunction with quantum-chemical calculations. The significant reactivity differences, favoring activation of the strong OH bond, can be ascribed to a proton-coupled electron transfer (PCET) mechanism operative in the activation of water. This study deepens our mechanistic understanding on how inert RH bonds are cleaved by metal oxides.

On the role of the electronic structure of the heteronuclear oxide cluster [Ga2Mg2O5 ](.+) in the thermal activation of methane and ethane: an unusual doping effect.

The reactivity of the heteronuclear oxide cluster [Ga2 Mg2 O5 ](.+) , bearing an unpaired electron at a bridging oxygen atom (Ob (.-) ), towards methane and ethane has been studied using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). Hydrogen-atom transfer (HAT) from both methane and ethane to the cluster ion is identified experimentally. The reaction mechanisms of these reactions are elucidated by state-of-the-art quantum chemical calculations. The roles of spin density and charge distributions in HAT processes, as revealed by theory, not only deepen our mechanistic understanding of CH bond activation but also provide important guidance for the rational design of catalysts by pointing to the particular role of doping effects.

Thermal ethane activation by bare [V2O5]⁺ and [Nb2O5]⁺ cluster cations: on the origin of their different reactivities.

The gas-phase reactivity of [V2O5](+) and [Nb2O5](+) towards ethane has been investigated by means of mass spectrometry and density functional theory (DFT) calculations. The two metal oxides give rise to the formation of quite different reaction products; for example, the direct room-temperature conversions C2H6→C2H5OH or C2H6→CH3CHO are brought about solely by [V2O5](+). In distinct contrast, for the couple [Nb2O5](+)/C2H6, one observes only single and double hydrogen-atom abstraction from the hydrocarbon. DFT calculations reveal that different modes of attack in the initial phase of C-H bond activation together with quite different bond-dissociation energies of the M-O bonds cause the rather varying reactivities of [V2O5](+) and [Nb2O5](+) towards ethane. The gas-phase generation of acetaldehyde from ethane by bare [V2O5](+) may provide mechanistic insight in the related vanadium-catalyzed large-scale process.

Reactivity of oxygen radical anions bound to scandia nanoparticles in the gas phase: C-H bond activation.

The activation of C-H bonds in alkanes is currently a hot research topic in chemistry. The atomic oxygen radical anion (O(-·)) is an important species in C-H activation. The mechanistic details of C-H activation by O(-·) radicals can be well understood by studying the reactions between O(-·) containing transition metal oxide clusters and alkanes. Here the reactivity of scandium oxide cluster anions toward n-butane was studied by using a high-resolution time-of-flight mass spectrometer coupled with a fast flow reactor. Hydrogen atom abstraction (HAA) from n-butane by (Sc2O3)(N)O(-) (N=1-18) clusters was observed. The reactivity of (Sc2O3)(N)O(-) (N=1-18) clusters is significantly sizedependent and the highest reactivity was observed for N=4 (Sc8O13(-)) and 12 (Sc24O37(-)). Larger (Sc2O3)(N)O(-) clusters generally have higher reactivity than the smaller ones. Density functional theory calculations were performed to interpret the reactivity of (Sc2O3)(N)O(-) (N=1-5) clusters, which were found to contain the O(-·) radicals as the active sites. The local charge environment around the O(-·) radicals was demonstrated to control the experimentally observed size-dependent reactivity. This work is among the first to report HAA reactivity of cluster anions with dimensions up to nanosize toward alkane molecules. The anionic O(-·) containing scandium oxide clusters are found to be more reactive than the corresponding cationic ones in the C-H bond activation.