Dynamics ofLens culinaris agglutinin studied by red-edge excitation spectra and anisotropy measurements of 2-p-toluidinylnaphthalene-6-sulfonate (TNS) and of tryptophan residues

The fluorescence of 2-p-toluidinylnaphthalene-6-sulfonate bound toLens culinaris agglutinin and of the Trp residues of the protein was investigated. Red-edge excitation spectra and steady-state anisotropy as a function of temperature indicate that the TNS is bound rigidly. Red-edge excitation spectra, steady-state anisotropy as a function of sucrose and anisotropy decay experiments performed on Trp residues fluorescence prove that the internal fluorophore presents residual motion independent of the global rotation of the protein. Fluorescence anisotropy decay allows to calculate the rotational correlation time (351 ps) of this local motion. Quenching resolved emission anisotropy with iodide gives values equal to 0.257 and 0.112 for the anisotropies of the buried and the surface Trp residues, respectively. This result indicates that the Trp residues present at the surface of the protein have important local motions compared to those embedded in the protein matrix. The results obtained from TNS and Trp residues indicate that the agglutinin has different dynamic domains.     

Luminescence and anisotropy decays ofN-3-pyrene maleimide labeling IgG proteins and cells

The Py.M (N-3-Pyrene Maleimide) is a dye that covalently binds to reactive amino or sulfhycryl groups to give highly fluorescent protein conjugates. Measurements of luminescence lifetimes and anisotropy decays have been performed with a Phase and Modulation Fluorometer. Complexes of Py.M-antibody (IgG antimouse) and tumoral cells C6 labeled with Py.M have been investigated. The Py.M fluorescence in buffer solution and the protein and cells natural fluorescence have been checked. For Py.M-IgG and labeled cells, the fluorescence decays present interesting behaviours. The least-squares analysis of the experimental results on Py.M-IgG complex points out two lorentzian distributions centered at 74 ns and 11 ns, on the contrary, for the labeled cells, a discrete component at 100 ns and a lorentzian distribution centered at 5 ns are shown. In both systems a weak component lower than 1 ns is observed. The fluorescence decays, mainly the long lifetime one, are very sensitive to oxygen quenching, showing the high efficiency of O₂ quenching. For samples N₂ bubbled, the lifetime experimental resuits show a decrease of the oxygen accessibility from free probe in solution to Py.M-IgG complex and to labeled cells, compatible with a more compact packing of the probe binding site. The experimental results of anisotropy decays of degassed samples show for Py.M-IgG complexes a long rotation correlation time of about 200 ns at T=5°C, assigned to overall rotation of the protein, besides shorter correlation times attributable to inner protein motions. For labeled cells, the long rotation correlation time becomes of the order of 580 ns confirming a progressive increase of the stabilization of the binding site.     

Conformational differences of oxytocin and vasopressin as observed by fluorescence anisotropy decays and transient effects in collisional quenching of tyrosine fluorescence

We used gigahertz frequency-domain fluorometry to examine the tyrosyl fluorescence intensity and anisotropy decays of the single-tyrosine cyclic peptide hormones oxytocin and vasopressin. Acrylamide quenching and a distance-dependent quenching model for collisional quenching were used to evaluate the extent of tyrosyl exposure to the quencher and to provide increased resolution of the picosecond anisotropy decays. Analysis of the intensity decays using a lifetime distribution model shows different distributions for oxytocin and vasopressin. We found that the tyrosyl fluorescence of lysine-vasopressin, as revealed both by the lifetime Stern-Volmer plots and from the quenching analysis, is quenched more effectively than oxytocin. ForN-acetyltyrosinamide (NATyrA), oxytocin, and lysine-vasopressin, we recovered apparent diffusion coefficients for quenching of 4.7×10^–6, 0.44×10^–6, and 4.3×10^–6 cm²/s, respectively, the lower value for oxytocin suggesting a shielded environment for its tyrosyl residue. Tyrosyl anisotropy decays were recovered by global analysis of progressively quenched samples. Compared with oxytocin, vasopressin displayed a longer correlation time for overall rotational diffusion and a higher amplitude for picosecond segmented motions of its tyrosyl residue. All the data are consistent with a more extended and flexible solution structure for vasopressin than for oxytocin.Dedicated to Professor Alfons Kawski on the occasion of his 65th birthday.     

The Conformational Changes of the Fungal Lipase from Humicola lanuginosa Induced by the Formation of Bile Salt and Mixed Micelles

The effect of sodium taurodeoxycholate and a mixture of sodium taurodeoxycholate/phosphatidyl-choline on the activation of fungal lipase from Humicola lanuginosa (HLL) was investigated by monitoring the specific activity and the changes in intrinsic protein fluorescence. A large increase in the inactivation rate was observed at about the critical micellar concentration of bile salt. On the contrary, a high activation of lipase was achieved by the presence of mixed micelles. Steady-state fluorescence quenching measurements were performed to resolve the fluorescence contribution of W89 residue in emission of four-tryptophan-containing HLL lipase. The W89 residue is located in the lid helix which participates in interfacial activation of the enzyme. The assignment of W89 residue was confirmed by use of the W89F mutant and inhibited form of lipase. The FQRS (fluorescence quenching resolved spectra) method was used to decompose the total emission spectrum of HLL lipase. The FQRS results show that the fluorescence of the W89 residue is similar in inhibited and inactivated HLL lipase and exhibits a maximum of emission at about 345 ± 1 nm (_ex = 295 nm). In the mixed micelle solution the fluorescence of the W89 residue may be resolved into two components, with fluorescence maxima at 337 and 347 nm, respectively (_ex = 295 nm). It is concluded that HLL lipase undergoes a conformational transition upon specific interactions with both anionic and mixed micelles, resulting in a change in the microenvironment of the W89 residue.     

Distance-dependent fluorescence quenching ofN-acetyl-l-tryptophanamide by acrylamide

We examined the time-dependent intensity decays ofN-acetyl-l-tryptophanamide (NATA) when collisionally quenched by acrylamide in propylene glycol over a range of temperatures. The intensity decays of NATA became increasingly heterogeneous in the presence of acrylamide. The NATA intensity decays were not consistent with the Collins-Kimball radiation boundary condition (RBC) model for quenching. The steady-state Stern-Volmer plots show significant upward curvature, and quenching of NATA by acrylamide was observed even in vitrified propylene glycol, where translational diffusion cannot occur during the lifetime of the excited state. These frequencydomain and steady-state data indicate a through-space quenching interaction between NATA and acrylamide, and the results are consistent with a rate constant for quenching that depends exponentially on the fluorophore-quencher separation distance. The exponential distance-dependent rate of quenching also explains the upward curvature of the Stern-Volmer plot, and the steady-state data aid in determining the interaction distance between NATA and acrylamide. These results suggest that the distance-dependent quenching rates need to be considered in the interpretation of acrylamide quenching of proteins.     

Limiting Dynamics for Spherical Models of Spin Glasses at High Temperature

We analyze the coupled non-linear integro-differential equations whose solution is the thermodynamical limit of the empirical correlation and response functions in the Langevin dynamics for spherical p-spin disordered mean-field models. We provide a mathematically rigorous derivation of their FDT solution (for the high temperature regime) and of certain key properties of this solution, which are in agreement with earlier derivations based on physical grounds. AMS (2000) Subject Classification: Primary: 82C44 Secondary: 82C31, 60H10, 60F15, 60K35     

Limiting Dynamics for Spherical Models of Spin Glasses at High Temperature

We analyze the coupled non-linear integro-differential equations whose solution is the thermodynamical limit of the empirical correlation and response functions in the Langevin dynamics for spherical p−spin disordered mean-field models. We provide a mathematically rigorous derivation of their FDT solution (for the high temperature regime) and of certain key properties of this solution, which are in agreement with earlier derivations based on physical grounds. AMS (2000) Subject Classification: Primary: 82C44, Secondery: 82C31, 60H10, 60F15, 60K35     

Determination of the in-plane anisotropy field in hexagonal systems via rotational magnetization: Theoretical model and Monte Carlo simulations

The magnetic anisotropy field in thin films with in-plane uniaxial anisotropy can be deduced from the VSM magnetization curves measured in magnetic fields of constant magnitudes. This offers a new possibility of applying rotational magnetization curves to determine the first- and second-order anisotropy constant in these films. In this paper we report a theoretical derivation of rotational magnetization curve in hexagonal crystal system with easy-plane anisotropy based on the principle of the minimum total energy. This model is applied to calculate and analyze the rotational magnetization process for magnetic spherical particles with hexagonal easy-plane anisotropy when rotating the external magnetic field in the basal plane. The theoretical calculations are consistent with Monte Carlo simulation results. It is found that to well reproduce experimental curves, the effect of coercive force on the magnetization reversal process should be fully considered when the intensity of the external field is much weaker than that of the anisotropy field. Our research proves that the rotational magnetization curve from VSM measurement provides an effective access to analyze the in-plane anisotropy constant K ₃ in hexagonal compounds, and the suitable experimental condition to measure K ₃ is met when the ratio of the magnitude of the external field to that of the anisotropy field is around 0.2. Supported by the National Natural Science Foundation of China (Grant Nos. 90505007 and 10774061) Recommended by LI FaShen     

Deuteron size effects on its elastic scattering from1H and4He

The effect of change in deuteron size on its elastic scattering from protons and alphas is investigated by varying the Hulthen parameters of the deuteron wave function in the scattering process. The cross sections forp-d scattering, calculated in the Born approximation, are found to increase substantially at backward angles even when the deuteron size is reduced by a small amount, whereas the shape of the angular distribution does not change significantly. For theα-d elastic scattering, interaction potential is obtained by folding the deuteron wave function and the optical potential for nucleon-scattering. The cross sections calculated atE _d = 13·7 MeV, shows that the first minimum around Θ_cm = 60° is deepend as the deuteron size is reduced, while at 52 MeV bombarding energy, the size effects are not very distinct. These observations are useful in the interpretation of deuteron cluster knockout reactions.     

Formation and dynamics of Saturn and its disk simulated by using a new N-body model

The formation of Saturn and its disk is simulated using a new N-body self-gravitational model. It is demonstrated that the formation of the disk and the planet is the result of gravitational contraction of a slowly rotated particle cloud that have a shape of slightly deformed sphere. The sphere was flattened by a coefficient of 0.8 along the axis of rotation. During the gravitational contraction, the major part of the cloud transformed into a planet and a minor part transformed into a disk. The thin structured disk is a result of the electromagnetic interaction in which the magnetic forces acting on charged particles of the cloud originate in the core of the planet. The simulation program gives such parameters of Saturn as the escape velocity of about 35 km/s at the surface, density, rotational velocities of the rings and temperature distribution.     

Dielectric properties and specific conductivity of armenian natural clinoptilolite irradiated by electrons

The results of dielectric properties and direct current specific electric conductivity measurements in Armenian natural clinoptilolite samples are presented. Electron irradiation with energy 8 MeV and thermal treatment of samples are performed to elucidate possible enhancement mechanisms of clinoptilolite parameters. The results are discussed on the basis of new point structural defects formation and recombination of initial ones in samples. It was shown that the irradiation dose of 3·10¹⁶ el/cm² is critical for natural zeolite structural change, which is manifested by significant changes in dielectric properties and other characteristics. The samples subjected to high temperature heating after electron irradiation, in comparison with an unquenched one, have a significantly higher (about an order of magnitude) value of specific conductivity.