Dynamic characteristic of amitriptyline in water by ultrasonic relaxation method and molecular orbital calculation.

Ultrasonic absorption coefficients in the frequency range of 0.8-220 MHz have been measured in aqueous solution of amitriptyline (3-(10,11-dihydro-5H-dibenzo[a,d]cycloheptene-5-ylidene)-N,N-dimethyl-1-propanamine) in the concentration range from 0.20 to 0.60 mol dm(-3) at 25 °C. A single relaxational phenomenon has been observed, and the relaxation frequency is independent of the concentration. It has been also observed that the amplitude of the relaxational absorption increases linearly with the analytical concentration. From these ultrasonic relaxation data, it has been concluded that the relaxation is associated with a unimolecular reaction due to a conformational change of the solute molecule, such as a structural change due to a rotational motion of a group in the solute molecule. Molecular orbital semiempirical methods using AM1 (Austin model 1) and PM3 (modified neglect of diatomic overlap parametric method 3) have been applied to obtain the standard enthalpy of formation for amitriptyline molecule at various dihedral angles around one of the bonds in alkylamine side chain. The results have shown the two clear minimum standard enthalpies of formation for amitriptyline. From the difference of the two values, the standard enthalpy change between the two stable conformers has been calculated be 2.9 kJ mol(-1). On a rough assumption that the standard enthalpy change reflects the standard free energy change, the equilibrium constant for the rotational isomers has been estimated to be 0.31. Combining this value with the experimental ultrasonic relaxation frequency, the backward and forward rate constants have been evaluated. The standard enthalpy change of the reaction has been also estimated from the concentration dependence of the maximum absorption per wavelength, and it has been close to that calculated by the semiempirical methods. The ultrasonic absorption measurements have been also carried out in amitriptyline solution in the presence of ß-cyclodextrin. However, the ultrasonic relaxation has not been found in the above frequency range. The result has been discussed in relation to the host-guest complex formation between ß-cyclodextrin and amitriptyline.

Role of the aromatic residues in the near-amino terminal motif of vimentin in intermediate filament assembly in vitro.

Type III and IV intermediate filament (IF) proteins share a conserved sequence motif of -Tyr-Arg-Arg-X-Phe- at the near-amino termini. To characterize significance of the aromatic residues in the motif, we prepared vimentin mutants in which Tyr-10 and Phe-14 are substituted with Asn and Ser (Vim[Y10N], Vim[F14S] and Vim[Y10N, F14S]), and examined assembly properties in vitro by electron microscopy and viscosity measurements. At 2 s after initiation of assembly reaction at pH 7.2 and 150 mM NaCl, all the vimentin mutants formed so-called unit-length filaments (ULFs) that were slightly larger than ULFs of wild-type vimentin. In following filament elongation, Vim[Y10N, F14S] and Vim[Y10N] performed longitudinal annealing of ULFs very rapidly and formed IFs within only 2.5 and 5 min, respectively, while Vim[F14S] and wild-type vimentin gave IFs by 40-60 min. The IFs of Vim[Y10N, F14S] and Vim[Y10N], however, tended to intertwine each other and formed bundles in parts of the specimens. The intertwinements decreased as the salt concentration decreased, and optimal salt concentration for the two mutants to form normal IFs was 50 mM. These results suggest that the aromatic residues, especially Tyr-10, in the motif have a role in controlling intermolecular interactions involved in IF assembly in vitro and suppress undesirable filament intertwinements at physiological ionic strength.

Inclusion kinetics of a nucleotide into a cyclodextrin cavity by means of ultrasonic relaxation.

To examine a dynamic interaction between nucleotide and cyclic oligosaccharide, ultrasonic absorption measurements were carried out in aqueous solution containing beta-cyclodextrin (beta-CD) and adenosine 5'-monophosphate (AMP) in the frequency range of 0.8-95 MHz. A relaxational absorption was observed in the solution, although it was not found in the individual solution of beta-CD or AMP. From the concentration dependences of AMP on the relaxation time and the maximum absorption per wavelength, the cause of the relaxation was attributed to a perturbation of a chemical equilibrium associated with a complex formation between beta-CD (host) and AMP (guest). The rate constants for the formation and breakup processes of the complex were determined. Also, a standard volume change of the reaction was obtained. From comparisons of the obtained rate and thermodynamic parameters with those for beta-CD and various guests, it has been concluded that the adenine moiety is included in the beta-CD cavity and that the hydrogen bonds may play a role in the complex formation.

Kinetic study for the inclusion complex of carboxylic acids with cyclodextrin by the ultrasonic relaxation method.

Ultrasonic absorption coefficients in the frequency range of 0.8-95 MHz were measured in aqueous solutions containing both beta-cyclodextrin (beta-CD) (host) and butanoic acid (in its dissociated form and undissociated one) (guest). A single relaxational phenomenon was observed only when the solutes were coexisting, although no relaxation was found in the beta-CD solution or in the acid solutions. The absorption was also measured in a solution of pentanoic acid (dissociated form) with beta-CD, and single relaxation was detected. The ultrasonic relaxation observed in these solutions was due to a perturbation of a chemical equilibrium related to a reaction of an inclusion complex formed by the host and guest. The equilibrium constant was obtained from the dependence of the maximum absorption per wavelength on the guest concentration. The rate constant for the inclusion process of the guest into a cavity of beta-CD and that for the leaving process from the cavity were determined from the obtained relaxation frequency and the equilibrium constant. The standard volume change of the reaction was also computed from the maximum absorption per wavelength. These results were compared with those in solutions containing both beta-CD and different guest molecules. It was found that the hydrophobicity of guest molecules played an important role in the formation of the inclusion complex and also that the charge on the carboxylic group had a considerable effect on the kinetic characteristics of the complexation reaction.

Dynamic study of interaction between beta-cyclodextrin and aspirin by the ultrasonic relaxation method.

A single ultrasonic relaxational phenomenon was observed in aqueous solutions containing both beta-cyclodextrin (beta-CD) as host and nonionized or ionized acetylsalicylic acid (aspirin) as guest. The observed relaxation was responsible for a dynamic complexation reaction between beta-CD and aspirin molecules, concomitant with a volume change during the reaction. The kinetic and equilibrium constants for the complexation in the acid (nonionized) form of the aspirin system were derived from the guest concentration dependence of the relaxation frequency. The equilibrium constant for the carboxylate (ionized) form of aspirin was determined from the concentration dependence of a maximum absorption per wavelength, and the rate constants were calculated by using the determined equilibrium constant and the observed relaxation frequencies, which remained nearly almost constant over the concentration range studied. The results showed that the effect of charge on the aspirin molecule was reflected only in the dissociation process from the beta-CD cavity, while no remarkable change was seen in the association process whose rate was diffusion controlled. The results could be explained on the basis of the difference of the hydrophobic moieties in the two guests that were included in the host cavity. The results of the standard volume change for the complexation reaction were closely related to the number of expelled water molecules originally located in the beta-CD cavity and the volume of the aspirin molecule incorporated into the beta-CD cavity.

Ultrasonic relaxation measurements in aqueous solution and molecular orbital calculation on imipramine.

Ultrasonic absorption coefficients have been measured in aqueous solution of imipramine {3-(10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)-N,N-dimethylpropan-1-amine} in the frequency range of 0.8-220 MHz at 25 °C. The frequency dependences of the observed absorption was characterized by a Debye-type relaxational equation with two relaxation frequencies, although only one relaxation had been observed in aqueous solutions of the related molecule amitriptyline. Both of the relaxation frequencies in imipramine solutions were found to be independent of the solute concentration and the amplitudes of the relaxational absorptions increase linearly with increasing solute concentration. It was therefore concluded that these two relaxations are associated with unimolecular reactions, such as a structural change due to rotational motions of the bond in the specified group in the imipramine molecule. To analyze quantitatively the source of the relaxations, semiempirical molecular orbital methods have been applied to determine the standard enthalpy of formation of the imipramine molecule at various dihedral angles around the bonds in the alkylamine side chain. According to the results, only one rotational motion of carbon-carbon bond in the side chain was found to be appropriate and the three minima of the standard enthalpy of formation was obtained as a function of the rotational angle. At the three minimum positions, the values of the standard enthalpy of formation are almost the same. With the assumptions (a) that rotational motion is not accompanied by a volume change of the reaction and (b) that the standard free energy change is close to the difference in the values between the standard enthalpies of formation, the equilibrium constants for the rotational isomerization have been calculated to be near unity. Hence, the forward and backward rate constants of the isomerization reactions are nearly the same. If one assumes that there are two kinds of rotational motions in one bond of the molecule, one proceeds with a rate constant on the order of 10(8) s(-1), whereas the other with a rate constant on the order of 10(6) s(-1). The faster and slower processes are also distinguished by the height of the standard enthalpy of formation.

Dynamic interaction between alkylammonium ions and beta-cyclodextrin by means of ultrasonic relaxation.

Ultrasonic absorption measurements in the frequency range from 0.8 to 220 MHz were carried out in aqueous solutions of pentylammonium chloride (PEACL) and hexylammonium chloride (HEACL) with beta-cyclodextrin (beta-CD) at pH approximately 7.2 and 25 degrees C. A single relaxational absorption was attributed to a perturbation of a chemical relaxation associated with the formation of a complex between beta-CD and the alkylammonium chlorides. The rate and equilibrium constants for the complexation reaction were determined from the concentration dependence of the relaxation frequency. Increasing the chain length of the alkylammonium ion led to an increase in the stability of the complex and slowed the exit rate of the ion from the beta-CD cavity. The standard volume change of the reaction was obtained from a maximum absorption per wavelength and was attributed to water molecules being expelled from the cavity with concomitant alkylammonium ion insertion.

Ultrasonic relaxation due to inclusion complex of amino acid by beta-cyclodextrin in aqueous solution.

Ultrasonic absorption coefficients in the frequency range of 0.8-95 MHz were measured in aqueous solutions of L-methionine or L-norleucine (guest) in the presence and absence of beta-cyclodextrin (beta-CD, host) at 25 degrees C. A single relaxational absorption was observed only in the solutions containing the guest and host. The ultrasonic relaxation was attributed to a perturbation of a chemical equilibrium associated with an interaction between beta-CD and the amino acid to form the host-guest complex. The kinetic and thermodynamic parameters in the system of L-norleucine with beta-CD were determined from the concentration dependence of the relaxation frequency and the maximum absorption per wavelength. Because of the concentration independence of the relaxation frequency in L-methionine system with beta-CD, the equilibrium constant and the standard volume change of the complexation reaction were estimated first from the concentration dependence of maximum absorption per wavelength, and subsequently the rate constants were calculated with the help of the estimated equilibrium constant and the observed relaxation frequency. The results obtained in this study were compared with those for systems of beta-CD with other amino acids or alcohols having comparable hydrophobicity.