Energetics of carbohydrate binding by a 14 kDa S-type mammalian lectin

The thermodynamics of the binding of derivatives of galactose and lactose to a 14 kDa beta-galactoside-binding lectin (L-14) from sheep spleen has been studied in 10 nM phosphate/150 mM NaCl/10 mM beta-mercaptoethanol buffer, pH 7.4, and in the temperature range 285-300 K using titration calorimetry. The single-site binding constants of various sugars for the lectin were in the following order: N-acetyl-lactosamine thiodigalactoside > 4-methylumbelliferyl lactoside > lactose > 4-methylumbelliferyl alpha-D-galactoside > methyl-alpha-galactose > methyl-beta-galactose. Reactions were essentially enthalpically driven with the binding enthalpies ranging from -53.8 kJ/mol for thiodigalactoside at 301 K to -2.2 kJ/mol for galactose at 300 K, indicating that hydrogen-bonding and van der Waals interactions provide the major stabilization for these reactions. However, the binding of 4-methylumbelliferyl-alpha-D-galactose displays relatively favourable entropic contributions, indicating the existence of a non-polar site adjacent to the galactose-binding subsite. From the increments in the enthalpies for the binding of lactose, N-acetyl-lactosamine and thiodigalactoside relative to methyl-beta-galactose, the contribution of glucose binding in the subsite adjacent to that for galactose shows that glucose makes a major contribution to the stability of L-14 disaccharide complexes. Observation of enthalpy-entropy compensation for the recognition of saccharides such as lactose by L-14 and the absence of it for monosaccharides such as galactose, together with the lack of appreciable changes in the heat capacity (delta Cp), indicate that reorganization of water plays an important role in these reactions.

Thermodynamics of monosaccharide binding to concanavalin A, pea (Pisum sativum) lectin, and lentil (Lens culinaris) lectin

Titration calorimetry measurements of the binding of methyl alpha-D-mannopyranoside (Me alpha Man), D-mannopyranoside (Man), methyl alpha-D-glucopyranoside (Me alpha Glu), and D-glucopyranoside (Glu) to concanavalin A (Con A), pea lectin, and lentil lectin were performed at 281 and 292 K in 0.01 M dimethylglutaric acid-NaOH buffer (pH 6.9) containing 0.15 M NaCl and Mn+2 and Ca+2 ions. The site binding enthalpies, delta H, are the same at both temperatures and range from -28.4 +/- 0.9 (Me alpha Man) to -16.6 +/- 0.5 kJ mol-1 (Glu) for Con A, from -26.2 +/- 1.1 (Me alpha Man) to -12.8 +/- 0.4 kJ mol-1 (Me alpha Glu) for pea lectin, and from -16.6 +/- 0.7 (Me alpha Man) to -8.0 +/- 0.2 kJ mol-1 (Me alpha Glu) for lentil lectin. The site binding constants range from 17 +/- 1 x 10(3) M-1 (Me alpha Man to Con A at 281.2 K) to 230 +/- 20 M-1 (Glu to lentil lectin at 292.6 K) and exhibit high specificity for Con A where they are in the Me alpha Man:Man:Me alpha Glu:Glu ratio of 21:4:5:1, while the corresponding ratio is 5:2:1.5:1 for pea lectin and 4:2:2:1 for lentil lectin. The higher specificity for Con A indicates more interactions between the amino acid residues at the binding site and the carbohydrate ligand than for the pea and lentil lectin-carbohydrate complexes. The carbohydrate-lectin binding results exhibit enthalpy-entropy compensation in that delta Hb (kJ mol-1) = -1.67 +/- 0.06 x 10(4) + (1.30 +/- 0.12)T(K) delta Sb (J mol-1K-1). Differential scanning calorimetry measurements on the thermal denaturation of the lectins and their carbohydrate complexes show that the Con A tetramer dissociates into monomers, while the pea and lentil lectin dimers dissociate into two submonomer fragments. At the denaturation temperature, one carbohydrate binds to each monomer of Con A and the pea and lentil lectins. Complexation with the carbohydrate increases the denaturation temperature of the lectin and the magnitude of the increases yield binding constants in agreement with the determinations from titration calorimetry.

Thermodynamics of the binding of galactopyranoside derivatives to the basic lectin from winged bean (Psophocarpus tetrogonolobus)

The thermodynamics of the binding of D-galactopyranoside (Gal), 2-acetamido-2-deoxygalactopyranoside (GalNAc), methyl-alpha-D-galactopyranoside, and methyl-beta-D-galactopyranoside to the basic agglutinin from winged bean (WBAI) in 0.02 M sodium phosphate and 0.15 M sodium chloride buffer have been investigated from 298.15 to 333.15 K by titration calorimetry and at the denaturation temperature by differential scanning calorimetry (DSC). WBAI is a dimer with two binding sites. The titration calorimetry yielded single-site binding constants ranging from 0.56 +/- 0.14 x 10(3) M-1 for Gal at 323.15 K to 7.2 +/- 0.5 x 10(3) M-1 for GalNAc at 298.15 K and binding enthalpies ranging from -28.0 +/- 2.0 kJ mol-1 for GalNAc at 298.15 K to -14.3 +/- 0.1 kJ mol-1 for methyl-beta-D-galactopyranoside at 322.65 K. The denaturation transition consisted of two overlapping peaks over the pH range 5.6-7.4. Fits of the differential scanning calorimetry data to a two-state transition model showed that the low temperature transition (341.6 +/- 0.4 K at pH 7.4) consisted of two domains unfolding as a single entity while the higher temperature transition (347.8 +/- 0.6 K at pH 7.4) is of the remaining WBAI dimer unfolding into two monomers. Both transitions shift to higher temperatures and higher calorimetric enthalpies with increase in added ligand concentration at pH 7.4. Analysis of the temperature increase as a function of added ligand concentration suggests that one ligand binds to the two domains unfolding at 341.6 +/- 0.6 K and one ligand binds to the domain unfolding at 347.8 +/- 0.6 K.

The proton magnetic resonance study of the structure of Na2Zn(SO4)2·4H2O

The proton magnetic resonance spectra of single crystals of Na2Zn(SO4)2·4H2O have been investigated and the orientations of the water molecules have been determined. Using the heavy atom structure determined by X-rays a system of hydrogen bonds between water and sulphate oxygens has been proposed.

Proton magnetic resonance study of the structure of two tutton's salts

Proton magnetic resonance spectra of single crystals of two Tutton's salts, K2Zn (SO4)2.6H22O and K2Mg (SO4)2.6H2O, have been studied and the orientations of the water molecules in the structure have been determined. Using the heavy-atom structure of (NH4) 2Mgt(SO4)2.6H2O as determined by x-ray diffraction, a system of hydrogen bonds between the water and sulfate oxygens in Tutton's salts has been proposed. It appears that the x-ray structure needs considerable refinement.

Determination of distances of sugar protons from Mn2+ in concanavalin A

1H NMR spin-lattice relaxation time (T1) measurements have been carried out with various sugars, viz. methyl alpha-D-glucopyranoside (alpha-MeGluP), methyl beta-D-lucopyranoside (beta-MeGluP), methyl alpha--annopyranoside (alpha-MeManP), maltose (4-O-alpha-D-glucopyranosyl--glucose), nigerose (3-O-alpha-D-glucopyranosyl-D-glucose), p-nitrophenyl alpha-maltoside (PNP-alpha-maltoside) and p-nitrophenyl beta-maltoside (PNP-beta-maltoside) to determine the distances of sugar protons from Mn2+ in concanavalin A (Con A). With a rotational correlation time of 1.58 x 10(-10) s determined, distances were calculated using Solomon-Bloembergen equation. The data obtained indicated differences in disposition of different groups in the binding site of Con A. An average value of about 10 A was obtained for the distances of sugar protons from Mn2+ in Con A. In the case of mono and disaccharides, the non-reducing end sugar unit was found to be closer to Mn2+ than the reducing end one.

Criticality of the exponential rate of decay for the largest nearest-neighbor link in random geometric graphs

Let n points be placed independently in d-dimensional space according to the density f(x) = A(d)e(-lambda parallel to x parallel to alpha), lambda, alpha > 0, x is an element of R-d, d >= 2. Let d(n) be the longest edge length of the nearest-neighbor graph on these points. We show that (lambda(-1) log n)(1-1/alpha) d(n) - b(n) converges weakly to the Gumbel distribution, where b(n) similar to ((d - 1)/lambda alpha) log log n. We also prove the following strong law for the normalized nearest-neighbor distance (d) over tilde (n) = (lambda(-1) log n)(1-1/alpha) d(n)/log log n: (d - 1)/alpha lambda <= lim inf(n ->infinity) (d) over tilde (n) <= lim sup(n ->infinity) (d) over tilde (n) <= d/alpha lambda almost surely. Thus, the exponential rate of decay alpha = 1 is critical, in the sense that, for alpha > 1, d(n) -> 0, whereas, for alpha <= 1, d(n) -> infinity almost surely as n -> infinity.

Modes of binding of α(1–2) linked manno-oligosaccharides to concanavalin A

Three-dimensional structures of the complexes of concanavalin A (ConA) with alpha(1-2) linked mannobiose, triose and tetraose have been generated with the X-ray crystal structure data on native ConA using the CCEM (contact criteria and energy minimization) method. All the constituting mannose residues of the oligosaccharide can reach the primary binding site of ConA (where methyl-alpha-D-mannopyranose binds). However, in all the energetically favoured complexes, either the non-reducing end or middle mannose residues of the oligosaccharide occupy the primary binding site. The middle mannose residues have marginally higher preference over the non-reducing end residue. The sugar binding site of ConA is extended and accommodates at least three alpha(1-2) linked mannose residues. Based on the present calculations two mechanisms have been proposed for the binding of alpha(1-2) linked mannotriose and tetraose to ConA.

Structure of basic winged-bean lectin and a comparison with its saccharide-bound form

The crystal structure of the saccharide-free form of the basic form of winged-bean agglutinin (WBAI) has been solved by the molecular-replacement method and refined at 2.3 Angstrom resolution The final R factor is 19.74b for all data in the resolution range 8.0-2.3 Angstrom. The asymmetric unit contains two half-dimers, each located on a crystallographic twofold axis. The structure of the saccharide-free form is compared with that of the complex of WBAI wi th methyl-alpha-D-galactoside. The complex is composed of two dimers in the asymmetric unit. The intersubunit interactions in the dimer are nearly identical in the two structures The binding site of the saccharide-free structure contains three ordered water molecules at positions similar to those of the hydroxyl groups of the carbohydrate which an hydrogen bonded to the protein. Superposition of the saccharide-binding sites of the two structures shows that the major changes involve expulsion of these ordered water molecules and a shift of about 0.6 Angstrom of the main-chain atoms of the variable loop.

Increased glycosidic bond stabilities in 4-C-hydroxymethyl linked disaccharides

Three new hydroxymethyl-linked non-natural disaccharide analogues, containing an additional methylene group in between the glycosidic linkage, were synthesized by utilizing 4-C-hydroxymethyl-alpha-D-glucopyranoside as the glycosyl donor. A kinetic study was undertaken to assess the hydrolytic stabilities of these new disaccharide analogues toward acid-catalyzed hydrolysis, at 60 degrees C and 70 degrees C. The studies showed that the disaccharide analogues were stable, by an order of magnitude, than naturally-occurring disaccharides, such as, cellobiose, lactose, and maltose. The first order rate constants were lower than that of methyl glycosides and the trend of hydrolysis rate constants followed that of naturally-occurring disaccharides. alpha-Anomer showed faster hydrolysis than the beta-anomer and the presence of axial hydroxyl group also led to faster hydrolysis among the disaccharide analogues. Energy minimized structures, derived through molecular modeling, showed that dihedral angles around the glycosidic bond in disaccharide analogues were nearly similar to that of naturally-occurring disaccharides. (C) 2011 Elsevier Ltd. All rights reserved.

New symmetrical dinucleating ligand based assembly of bridged dicopper(II) and dizinc(II) centers: Synthesis, structure, spectroscopy, magnetic properties and glycoside hydrolysis

Two dinuclear copper(II) complexes Li(H2O)(3)(CH3OH)](4)Cu2Br4]Cu-2(cpdp)(mu-O2CCH3)](4)(OH)(2) (1), Cu (H2O)(4)]Cu-2(cpdp)(mu-O2CC6H5)](2)Cl-2 center dot 5H(2)O (2), and a dinuclear zinc(II) complex Zn-2(cpdp)(mu-O2CCH3)] (3) have been synthesized using pyridine and benzoate functionality based new symmetrical dinucleating ligand, N, N'-Bis2-carboxybenzomethyl]-N, N'-Bis2-pyridylmethyl]-1,3-diaminopropan-2-ol (H(3)cpdp). Complexes 1, 2 and 3 have been synthesized by carrying out reaction of the ligand H3cpdp with stoichiometric amounts of Cu-2(O2CCH3)(4)(H2O)(2)], CuCl2 center dot 2H(2)O/C6H5COONa, and Zn(CH3COO)(2)center dot 2H(2)O, respectively, in methanol in the presence of NaOH at ambient temperature. Characterizations of the complexes have been done using various analytical techniques including single crystal X-ray structure determination. The X-ray crystal structure analyses reveal that the copper(II) ions in complexes 1 and 2 are in a distorted square pyramidal geometry with Cu-Cu separation of 3.455(8) angstrom and 3.492(1)angstrom, respectively. The DFT optimized structure of complex 3 indicates that two zinc(II) ions are in a distorted square pyramidal geometry with Zn-Zn separation of 3.492(8)angstrom. UV-Vis and mass spectrometric analyses of the complexes confirm their dimeric nature in solution. Furthermore, H-1 and C-13 NMR spectroscopic investigations authenticate the integrity of complex 3 in solution. Variable-temperature (2-300 K) magnetic susceptibility measurements show the presence of antiferromagnetic interactions between the copper centers, with J = -26.0 cm(-1) and -23.9 cm(-1) ((H) over cap = -2JS(1)S(2)) in complexes 1 and 2, respectively. In addition, glycosidase-like activity of the complexes has been investigated in aqueous solution at pH similar to 10.5 by UV-Vis spectrophotometric technique using p-nitrophenyl-alpha-D-glucopyranoside (4) and p-nitrophenyl-beta-D-glucopyranoside (5) as model substrates. (C) 2015 Elsevier B.V. All rights reserved.

The isotopic composition of energetic particles emitted from a large solar flare

We report measurements of isotope abundance ratios for 5-50 MeV/nuc nuclei from a large solar flare that occurred on September 23, 1978. The measurements were made by the Heavy Isotope Spectrometer Telescope (HIST) on the ISEE-3 satellite orbiting the Sun near an Earth-Sun libration point approximately one million miles sunward of the Earth. We report finite values for the isotope abundance ratios ¹³C/¹²C, ¹⁵N/¹⁴N, ¹⁸O/¹⁶O, ²²Ne/ ²⁰Ne, ²⁵Mg/²⁴Mg, and ²⁶Mg/²⁴Mg, and upper limits for the isotope abundance ratios ³He/⁴He, ¹⁴C/¹²C, ¹⁷O/¹⁶O, and ²¹Ne/²⁰Ne.

We measured element abundances and spectra to compare the September 23, 1978 flare with other flares reported in the literature. The flare is a typical large flare with "low" Fe/O abundance (≤ 0.1).

For ¹³C/¹²C, ¹⁵N/¹⁴N, ¹⁸O/¹⁶O, ²⁵Mg/ ²⁴Mg, and ²⁶Mg/²⁴Mg, our measured isotope abundance ratios agree with the solar system abundance ratios of Cameron (1981). For neon we measure ²²Ne/²⁰Ne = 0.109 + 0.026 - 0.019, a value that is different with confidence 97.5% from the abundance measured in the solar wind by Geiss at al. (1972) of ²²Ne/²⁰Ne = 0.073 ± 0.001. Our measurement for ²²Ne/²⁰Ne agrees with the isotopic composition of the meteoritic component neon-A.

Separate arguments appear to rule out simple mass fractionation in the solar wind and in our solar energetic particle measurements as the cause of the discrepancy in the comparison of the apparent compositions of these two sources of solar material.

Tm:Y_2SiO_5晶体的生长和光谱性质研究

采用中频感应提拉法生长了高质量的Tm:Y2SiO5(Tm:YSO)晶体,测定了晶体的晶格常数和分凝系数.运用劳厄照相法确定了单斜晶系Tm:YSO晶体的三个偏振轴〈010〉,D1和D2,在室温下测量了三个偏振轴方向的吸收光谱、荧光光谱和荧光寿命,计算了晶体吸收峰的吸收线宽和吸收截面.研究发现,相对于其他两个偏振轴方向,D1方向在790 nm处出现较强的吸收峰,同时在2μm附近出现了一定强度的发射峰,D1方向的吸收截面较大,荧光寿命较长.Tm:YSO晶体适用于AlGaAs二极管抽运固体激光器,在2μm波段固体激光器的应用上将有很大的发展潜力.

Enhanced photoresponse from the ordered microstructure of naphthalocyanine-carbon nanotube composite film

Naphthalocyanine-sensitized multi-walled carbon nanotube (NaPc-MWNT) composites have been synthesized through the pi-stacking between naphthalocyanine (NaPc) and carbon nanotubes. The resultant nanocomposites were characterized with a scanning electron microscope (SEM), a transmission electron microscope (TEM), and by UV - vis absorption and photocurrent spectra. The long-range ordering was observed in the NaPc - MWNT composites by using a TEM. The enhancement in the absorption intensity and the broadening of the absorption wavelength observed in the composite films, which were due to the attachment of NaPc on the MWNT surface, is discussed based on the measured UV - vis absorption spectra. Furthermore, the photoconductivity of the poly( 3-hexylthiophene)(PAT6) - NaPc - MWNT composite film was found to increase remarkably in the visible region and broaden towards the red regions. These new phenomena were ascribed to the larger donor/acceptor (D/A) interface and the formation of a biconsecutive D/A network structure, as discussed in consideration of the photoinduced charge transfer between PAT6 and NaPc - MWNT.

Dielectric relaxation of poled polymer DCNP/PMMA

We have prepared the polymer thin films of a 3-(1,1-dicyanothenyl)-1-phenyl-4,5-dihydro-1H-pyrazole (DChTP)/poly (methyl methacrylate) (PMMA) guest-host system by spin coating. In order to investigate their temporal and temperature stability, we have measured their dielectric relaxation spectra including the frequency dependence of the real and imaginary parts of dielectric constants. The investigated frequency ranged from 50 Hz to 10 MHz. The measured temperature range above the glass transition temperature T-g (95 degrees C) of the DCNP/PMMA system was from 95 degrees C to 1250C. By using the Adam-Gibbs model, the temperature dependence of the characteristic time tau above T-g was fitted, and the values of the characteristic times tau below T-g were estimated. The lifetimes of the polymer were evaluated by the Kohlrausch-Williams Watts (KWW) empirical decay model. (C) 2000 Elsevier Science B.V. All rights reserved.