The use of ECD/ETD to identify the site of electrostatic interaction in noncovalent complexes

Electrostatic interactions play an important role in the formation of noncovalent complexes. Our previous work has highlighted the role of certain amino acid residues, such as arginine, glutamate, aspartate, and phosphorylated/sulfated residues, in the formation of salt bridges resulting in noncovalent complexes between peptides. Tandem mass spectrometry (MS) studies of these complexes using collision-induced dissociation (CID) have provided information on their relative stability. However, product-ion spectra produced by CID have been unable to assign specifically the site of interaction for the complex. In this work, tandem MS experiments were conducted on noncovalent complexes using both electron capture dissociation (ECD) and electron-transfer dissociation (ETD). The resulting spectra were dominated by intramolecular fragments of the complex with the electrostatic interaction site intact. Based upon these data, we were able to assign the binding site for the peptides forming the noncovalent complex.     

Production and Characterization of an Alkaline Thermostable Crude Lipase from an Isolated Strain of <Emphasis Type="Italic">Bacillus cereus</Emphasis> C<Subscript>7</Subscript>

A bacterial strain isolated from spoiled coconut and identified as Bacillus cereus was found capable of producing alkaline thermostable extracellular lipase. Optimum temperature, time, and pH for enzyme substrate reaction were found to be 60 °C, 10 min, and 8.0 respectively. Common surfactants except Triton X 100 and cetyltrimethylammonium bromide have no or very little inhibitory effects on enzyme activity. The enzyme was found to be stable in presence of oxidizing agents and protease enzyme. The maximum lipase production was achieved at 30–33 °C, pH 8.0 on 24 h of fermentation using 50 ml medium in a 250-ml Erlenmeyer flask. The superior carbon and nitrogen sources for lipase production were starch (2%) and ammonium sulfate (nitrogen level 21.2 mg/100 ml), peptone (nitrogen level 297 mg/100 ml), and urea (nitrogen level 46.62 mg/100 ml) in combination, respectively. The maximum enzyme activity obtained was 33 ± 0.567 IU/ml.     

Syntheses, X-ray crystal structures, DNA binding, oxidative cleavage activities and antimicrobial studies of two Cu(II) hydrazone complexes

From a mononuclear Cu(II)-hydrazone complex [Cu(PBH)₂] (1), one μ_1,1-azido bridged dinuclear Cu(II) complex having the formula [{Cu(PBH)(μ_1,1-NNN)}₂] (2) (where HPBH = 2-pyridinecarboxaldehyde benzoyl hydrazone) has been synthesised. Both the complexes are characterised by elemental analyses, IR and UV–Vis spectroscopic studies. The tridentate hydrazone pro-ligand (HPBH) is obtained by the condensation of benzhydrazide and pyridine-2-carboxaldehyde. The structures of the complexes have conclusively been established by the X-ray single crystal diffraction method. Complex 1 and 2 both display DNA binding ability, which is ascertained by UV–Vis titration and cyclic voltammetric studies using calf thymus DNA (CT-DNA). The apparent binding constants (K_app) are of moderate values and are 2.048 × 10⁴ M⁻¹ (±0.006) and 1.644 × 10⁴ M⁻¹ (±0.005), respectively. The modes of binding of the complexes with CT-DNA has been investigated using circular dichroism, ethidium bromide displacement assay and viscosity measurements. The cleavage properties of these complexes as well as the free pro-ligand with super coiled (SC) pUC19 are studied using the gel electrophoresis method, where both the complexes displayed chemical nuclease activity in the presence of H₂O₂ via an oxidative mechanism. The antimicrobial study using the free pro-ligand, 1 and 2 against both Gram positive and Gram negative bacteria are performed, 2 showed antimicrobial activity against both Gram negative and Gram positive bacteria whereas the free ligand and 1 show no antibacterial activity.     

Development and Validation of a New Analytical Method for the Determination of Related Components in Quetiapine Hemifumarate

A simple, sensitive isocratic rapid resolution liquid chromatographic assay method has been developed for the quantitative determination of quetiapine hemifumarate in bulk active pharmaceutical ingredient, used for the treatment of schizophrenia. The developed method is also applicable for the process related impurities determination. Efficient chromatographic separation was achieved on a C18 stationary phase with simple mobile phase combination delivered in a isocratic mode and quantification was by ultraviolet detection at 225 nm at a flow rate of 1.0 mL min^?1. In the developed LC method the resolution between quetiapine hemifumarate and its three potential impurities was found to be greater than 2.0. Regression analysis showed an r value (correlation coefficient) greater than 0.99 for quetiapine hemifumarate and its three impurities. This method was capable to detect all three impurities of quetiapine hemifumarate at a level of 0.003% with respect to test concentration of 1.0 mg mL^?1 for a 3 μL injection volume. The bulk active pharmaceutical ingredient was subjected to stress conditions of hydrolysis, oxidation, photolysis and thermal degradation. Considerable degradation was found to occur in oxidative stress conditions. The stress samples were assayed against a qualified reference standard and the mass balance was found close to 99.5%. The developed RR-LC method was validated with respect to linearity, accuracy, precision and robustness.     

Control of the Supramolecular Excimer Formation of Thioflavin T within a Cucurbit[8]uril Host: A Fluorescence On/Off Mechanism

On or off? A new excimer band at λ=570 nm was visualized during the noncovalent host–guest interaction between thioflavin T (ThT) and cucurbit[8]uril (CB8). Controlled dissociation of this assembly in the presence of Ca²⁺ was demonstrated as an on/off fluorescence switch (see picture).

     

A fixed point result in Menger spaces using a real function

The main result of this paper is a fixed point theorem of self-mappings in Menger spaces which satisfy certain inequality. This inequality involves a class of real functions which we call Φ-functions. As a corollary we obtain a result in the corresponding metric spaces. The result is supported by an example. The class of real functions we have used is the conceptual extension of altering distance functions used in metric fixed point theory.      

Resonance Raman spectroscopic study of cation radicals of aromatic molecules formed in the interlayer of transition-metal exchanged montmorillonites

Resonance Raman spectroscopy has been demonstrated to give important structural information on the reactions of aromatic molecules in the interlayer of transition-metal ion-exchanged montmorillonites. Para-substituted benzenes or 4,4-substituted biphenyls are oxidized to form their cation radicals, which are stabilized in the interlayer of the clay mineral. The oxidative dimerization or polymerization results in the formation of biphenyl type cations and poly-p-phenylene cations from mono-substituted benzenes and benzene, respectively.     

A theoretical study of surface plasmon cross coupling in asymmetric corrugated metal films

We investigate numerically the transmission properties of a thin sinusoidally corrugated metal film bounded by two different dielectrics in the context of the experiment of Gruhlkeet al (1986). We study the dominant contributions from both the propagating and evanescent plane waves. A comparison with the experimental results reveals that the decisive role in cross coupling is played by the evanescent waves emitted by the molecular dipole. We extend our studies to different corrugation amplitudes and widths to show their effect on transmission.     

Spontaneous assembly of a polymeric helicate of sodium with LVO(2) units forming the strand: photoinduced transformation into a mixed-valence product

The anionic cis-dioxovanadium(V) complex species LVO(2)(-) of a tridentate ONS ligand (H(2)L) can bind sodium ion in a bis-monodentate fashion like a bridging carboxylate group. The product [LVO(2)Na(H(2)O)(2)](infinity) (1) is a water soluble polymeric compound in which the complementary units are held together by the simultaneous use of hydrogen bonding and Coulombic interactions. Crystallographic characterization reveals that 1 is a single stranded helicate with LVO(2)(-) units forming the strand which surrounds the labile sodium ions that occupy the positions on the axis. In solution of protic solvents, viz. water and methanol, 1 is quite stable as indicated by electrical conductivity and (1)H NMR measurements. In aprotic solvents, viz. CH(3)CN, DMF, or DMSO, however, the extended hydrogen bonded network in 1 breaks apart and the helical structure collapses when irradiated with visible light. The product is a mixed-oxidation vanadium(IV/V) species obtained by photoinduced reduction as confirmed by EPR, time dependent (1)H NMR, and electronic spectroscopy. Compound 1 is a rare example of a nonnatural helix where hydrogen bonding interactions play a crucial role in stabilizing the single stranded polymeric structure such as that frequently observed in the biological world.