UDP-GlcNAc:Ser-protein N-acetylglucosamine-1-phosphotransferase from Dictyostelium discoideum recognizes serine-containing peptides and eukaryotic cysteine proteinases

Phosphoglycosylation catalyzed by UDP-GlcNAc:Ser-protein N-acetylglucosamine-1-phosphotransferase (Ser:GlcNAc phosphotransferase) adds GlcNAcalpha-1-P to peptidyl-Ser of selected Dictyostelium discoideum proteins. Lysosomal cysteine proteinase (CP), proteinase-1(CP7), is the major phosphoglycosylated protein in bacterially grown amoebae. GlcNAc-1-P is added within a Ser-rich domain containing SSS, SGSG, or SGSQ repeated motifs that are not found in other papain-like CPs. We studied the substrate specificity of the transferase using peptides containing these motifs and 12 other peptides with one or more Ser residues. Phosphoglycosylation is comparable for all three Dictyostelium CP motifs, but it is not restricted to them. Flanking residues in the other peptides strongly influence phosphoglycosylation efficiency. Dictyostelium microsomal membranes also phosphoglycosylate endogenous acceptors, and some of these acceptors occur as an 18 S complex with the transferase. CP-serine motif peptides inhibit endogenous acceptor phosphoglycosylation weakly (30-40%) at 800 microM, whereas catalytically inactive proteinase-1(CP7) and other non-phosphoglycosylated eukaryotic CPs, lacking the serine domain, inhibit transferase activity at 1-4 microM. SDS denaturation destroys the inhibitory potential of all CPs showing that transferase recognizes a conformation-dependent feature that is shared by all. Proteinase-1(CP7) expressed in Escherichia coli lacks GlcNAc-1-P, but it is a substrate for Ser:GlcNAc phosphotransferase, Km = 5.6 microM. Thus, Ser:GlcNAc phosphotransferase recognizes both acceptor peptide sequences and a conformational feature of eukaryotic CPs. This may be physiologically important for establishing or maintaining non-overlapping groups of GlcNAc-1-P- and Man-6-P-modified Dictyostelium proteins that reside in functionally distinct endo-lysosomal vesicles.     

Inhibition by adenosine triphosphate of heart microsomal neutral lipase activity

Triacylglycerol lipolysis was inhibited by palmitate, in the isolated perfused normal rat heart. Acetate or acetylcarnitine could reproduce the inhibitory effects of palmitate. Since heart neutral lipase plays an important role in the lipolysis of heart triacylglycerols, the effects of acetylcarnitine, acetyl CoA and related metabolites on the microsomal neutral lipase activity were studied. ATP inhibited the enzyme activity in a concentration-dependent manner without a lag phase. AMP and adenylyl imidodiphosphate, two compounds structurally related to ATP but whose phosphate groups cannot be transferred, did not inhibit the microsomal lipase activity. These results suggested that ATP inhibited the lipase activity through the transfer of its phosphate group. It is proposed that cellular ATP concentration is a determinant of tricylglycerol lipolysis in the heart.     

Structure and properties of polyamide 6 and 4-aminomethylcyclohexane carboxylic acid copolymers with an unusually short helical pitch for nylons

Segments of 4-aminomethylcyclohexane carboxylic acid (AMCC) inserted into the polyamide 6 (nylon 6, N6) chain are distributed in both the crystalline lattice and the amorphous regions. Thermal, spectroscopic and X-ray diffraction data indicate that the N6/AMCC copolymer crystallizes in an otherwise thermodynamically unfavorable γ crystalline form of N6. The chain-axis repeat or the helical pitch of 15.7 Å of this copolymer is the shortest ever reported for N6. This is accompanied by an expansion of the lattice in the equatorial plane. This short pitch was observed at comonomer concentrations as low as 10 mol%, and the lattice remains otherwise essentially same as that of N6 up to about 30 mol% AMCC. Conformational constraints imposed by interchain hydrogen bonding between N6 and AMCC with chain-axis repeat distance of 17.2 and 15.8 Å, respectively, are thought to be the reason for this shortened helical pitch. This short pitch (length scale ∼10 Å) results in a lower chain-modulus. Differences in the H-bond strength are invoked to explain the higher glass transition temperature in the dry copolymer, i.e. the increased stiffness of the multiple chain segments (length scales >10 Å) and the differing response to guest molecules such as water. Although the mechanical properties (tensile strength, shrinkage and creep) of the dry copolymer and the homopolymer were similar, moisture had a dramatically different effect on the two polymers. This behavior is attributed to the differences in lamellar morphology (length scales ∼100 Å), especially the contribution of the amorphous regions, which determine the bulk properties.     

SOLID-STATE REACTION BETWEEN BARIUM SULFATE AND CARBON

Barium sulfate is the chief source for the manufacture of barium compounds, which have a wide range of applications in chemical industries. In the preparation of these compounds, the first step is the reaction between barium sulfate and carbon to produce barium sulfide. In the present work, the mechanism of reaction between barium sulfate and carbon is investigated experimentally, covering a wide range of temperatures, times of reaction, and stoichiometric ratios of reactants. The reaction is conducted in air, nitrogen, and vacuum. It is also studied using the reactants in pellet form to determine the effect of compaction pressure on rate of reaction. A model is developed to estimate the kinetic parameters and mechanism of the reaction. An attempt is also made to study the feasibility of conducting the reaction for continuous production of barium sulfide in a moving bed rector using the reactants in pellet form.     

Orientation and Grain Boundary Microstructure of Alumina in Al/Al2O3 Composites Produced by Reactive Metal Penetration

The orientation and grain boundary microstructure of alumina in reactive metal penetration Al/Al₂O₃ composites are studied using orientation imaging microscopy and the results are compared with those of sintered polycrystalline Al₂O₃. The interconnected Al₂O₃ in the composite material is separated by Σ3 boundaries (twins) with a 60° rotation around the [0001] direction. A high frequency (∼100%) of Σ3 coincidence boundaries in composite alumina is remarkable since only ∼12% of boundaries in a sintered polycrystalline Al₂O₃ are of special nature. The coincidence boundaries in the in situ alumina grow in a coherent and faceted manner.     

Lamotrigine‐loaded polyacrylate nanoparticles synthesized through emulsion polymerization

A series of copolymeric nanoparticles of the partially water‐soluble monomer ethyl methacrylate and the water‐soluble monomer 2‐hydroxyl ethyl methacrylate were synthesized from emulsions containing sodium dodecyl sulfate via free‐radical polymerization. Lamotrigine, as a model drug, was loaded in nanoparticles during in situ polymerization. A stable and transparent poly(ethyl methacrylate‐co‐hydroxyl ethyl methacrylate) nanolatex was produced for all compositions and characterized for particle size by dynamic light scattering and transmission electron microscopy. Particles were found to be smaller than 50 nm in size. Structural characterization of copolymers was done by infrared spectrometry, gel permeation chromatography, and NMR spectroscopy. Drug encapsulation efficiency was determined by ultraviolet (UV)–visible spectrometry and was found to be 26–62% for copolymers with different compositions. UV data suggest molecular‐level dispersion of the drug in the nanoparticles. In vitro drug‐release studies showed the controlled release of lamotrigine. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

CFD analysis of low temperature water gas shift reactor

In the design of water gas shift reactors, the performance of catalysts is not known a priori and hence having a general kinetic expression will be of much help. Computational Fluid Dynamic study was carried out to investigate the performance of a packed bed reactor for different feed compositions using five commonly used types of macro kinetic models. User Defined Functions were developed for the reaction rate to predict the CO conversion in the reactor. The effects of temperature and time factor on CO conversion were studied. The Langmuir-Hinshelwood model gave the best prediction for H₂ rich mixtures. The Temkin model was better for higher CO concentrations, whereas the other models gave large deviations for the fixed bed reactor.     

CFD analysis of water gas shift membrane reactor

Fuel cell based modular power generation can be achieved by miniaturization and process intensification of equipments in the process. Fuel cells require hydrogen rich gas which can be generated through reforming and water gas shift reaction. The water gas shift reactor being kinetically limited occupies more volume to achieve the required CO conversion. A membrane reactor integrates the reaction and hydrogen separation stages and hence reduces the volume requirement. Computational Fluid Dynamics offers virtual prototyping of the reactor and thus helps in design, optimization and scale up of reactors. In this study customized User Defined Functions (UDFs) were developed to analyze the performance of low temperature water gas shift membrane reactor. The models were validated using literature data for the parameters – synthesis gas compositions, time factor, sweep flow rate and steam to CO ratio. The effect of all these parameters on the reactor was analyzed for CO conversion, H₂ recovery, DaPe, concentration polarization, concentration profiles and conversion index. The simulations have showed that the UDFs developed were capable of simulating the membrane reactor and this can be used for the design and optimization of the membrane reactor for any process conditions.     

Revisiting the mechanism of the triosephosphate isomerase reaction: the role of the fully conserved glutamic acid 97 residue

An analysis of 503 available triosephosphate isomerase sequences revealed nine fully conserved residues. Of these, four residues—K12, H95, E97 and E165—are capable of proton transfer and are all arrayed around the dihydroxyacetone phosphate substrate in the three‐dimensional structure. Specific roles have been assigned to the residues K12, H95 and E165, but the nature of the involvement of E97 has not been established. Kinetic and structural characterization is reported for the E97Q and E97D mutants of Plasmodium falciparum triosephosphate isomerase (Pf TIM). A 4000‐fold reduction in k_cat is observed for E97Q, whereas the E97D mutant shows a 100‐fold reduction. The control mutant, E165A, which lacks the key catalytic base, shows an approximately 9000‐fold drop in activity. The integrity of the overall fold and stability of the dimeric structure have been demonstrated by biophysical studies. Crystal structures of E97Q and E97D mutants have been determined at 2.0 Å resolution. In the case of the isosteric replacement of glutamic acid by glutamine in the E97Q mutant a large conformational change for the critical K12 side chain is observed, corresponding to a trans‐to‐gauche transition about the Cγ? Cδ (χ³) bond. In the E97D mutant, the K12 side chain maintains the wild‐type orientation, but the hydrogen bond between K12 and D97 is lost. The results are interpreted as a direct role for E97 in the catalytic proton transfer cycle. The proposed mechanism eliminates the need to invoke the formation of the energetically unfavourable imidazolate anion at H95, a key feature of the classical mechanism.     

Single-Step Synthesis of Chemically Cross-Linked Polysilastyrene and Its Conversion to β-Silicon Carbide

A new method for chemically cross-linking polysilastyrene using divinylbenzene as the cross-linking agent is reported. The procedure involves a single-step synthesis using the alkali-metal sodium to promote the polymerization of dimethyldichlorsilane in the presence of the comonomers phenylmethyldichlorosilane and divinylbenzene. The cross-linked polymer can be readily converted to β-SiC on pyrolysis at 1500°C. The β-SiC obtained by this procedure is nanocrystalline and has a grain-size distribution of 8–20 nm.