Nearest-neighbour-interaction model in the coupled-optical-phonon-mode theory of the infrared dispersion in monoclinic crystals: Application to Tutton salt single crystal.

The coupled-optical-phonon-mode theory of Barker and Hopfield of two mode interaction in isotropic crystals has been extended to monoclinic crystals. The analytical expressions for the calculation of the dielectric tensor elements in the ac crystal plane have been derived. It has been shown that the interaction dielectric model is a generalized expression of the dielectric tensor for monoclinic case when no interaction between modes is present.

The perturbation theory model of a spherical oscillator in electric field and the vibrational stark effect in polyatomic molecular species.

The effect of external electrostatic fields on the spherical oscillator energy states was studied using stationary perturbation theory. Besides the spherical oscillator with ideal symmetry, also a variety of the deformed systems were considered in which the deformations may be induced by the external fields, but also by the short-range crystal lattice forces. The perturbation theory analysis was carried out using the field-dependent basis functions.

The C-terminal and beta-wing regions of ammodytoxin A, a neurotoxic phospholipase A2 from Vipera ammodytes ammodytes, are critical for binding to factor Xa and for anticoagulant effect.

Ammodytoxin A (AtxA) from the venom of Vipera ammodytes ammodytes belongs to group IIA secreted phospholipase A2 (sPLA2), for which the major pathologic activity is presynaptic neurotoxicity. We show here that this toxin also affects hemostasis because it exhibits strong anticoagulant activity. AtxA binds directly to human coagulation factor Xa (FXa) with Kdapp of 32 nM, thus inhibiting the activity of the prothrombinase complex with an IC50 of 20 nM.

Basic amino acid residues in the beta-structure region contribute, but not critically, to presynaptic neurotoxicity of ammodytoxin A.

The molecular mechanism of action of presynaptically toxic secreted phospholipases A2 (sPLA2s) isolated from snake venoms is not completely understood. It has been proposed that the positive charge in the beta-structure region is important for their toxic activity. To test this hypothesis, we characterised several mutants of ammodytoxin A (AtxA) possessing substitution of all five basic residues in this region.

Vibrational states of tetrahedral molecular species in electric fields. A theoretical model for quantitative description of the latent symmetry effects in vibrational spectroscopy of matrix isolated tetrahedral species.

A theoretical model that enables a quantitative description of latent symmetry effects in the case of matrix isolated tetrahedral species is developed. Several particular cases are considered, that are of special interest from vibrational spectroscopic viewpoint. It is shown that the observed appearances of the IR spectra of various XY4n- anions isomorphously isolated in solid matrices may be systematically explained in terms of the orientation of the effective local crystalline field vector.

Identification of a novel binding site for calmodulin in ammodytoxin A, a neurotoxic group IIA phospholipase A2.

The molecular mechanism of the presynaptic neurotoxicity of snake venom phospholipases A2 (PLA2s) is not yet fully elucidated. Recently, new high-affinity binding proteins for PLA2 toxins have been discovered, including the important intracellular Ca2+ sensor, calmodulin (CaM). In the present study, the mode of interaction of group IIA PLA2s with the Ca2+-bound form of CaM was investigated by mutational analysis of ammodytoxin A (AtxA) from the long-nosed viper (Vipera ammodytes ammodytes).

mRNA secondary structure can greatly affect production of recombinant phospholipase A(2) toxins in bacteria.

The neurotoxic activity of ammodytoxin A (AtxA), a phospholipase A(2) from Vipera ammodytes ammodytes venom, has been investigated by protein engineering. With the aim of obtaining AtxA as a non-fused protein in the bacterial cytoplasm and avoiding problems with incomplete cleavage in vivo of the initial Met preceding the first residue (Ser1), a double mutant (S1A/E4Q) was prepared and expressed in Escherichia coli. Immunoblotting of the bacterial lysate showed that the mutant was synthesized at a low level not exceeding 0.

The amino acid region 115-119 of ammodytoxins plays an important role in neurotoxicity.

Quadruple (Y115K/I116K/R118M/N119L) and double (Y115K/I116K) mutants of ammodytoxin A, a presynaptically toxic phospholipase A(2) from Vipera ammodytes ammodytes venom, were prepared and characterized. The enzymatic activity of the quadruple mutant on phosphatidylcholine micelles was threefold higher than that of AtxA, presumably due to higher phospholipid-binding affinity, whereas the activity of the double mutant was twofold lower. The substantial decrease by more than two orders of magnitude in the lethal potency of both mutants, together with their decreased binding affinity for neuronal receptors, indicates involvement of the amino acid region 115-119 in neurotoxicity.

Tissue expression and immunolocalization of a novel human cathepsin P.

The mRNA of a novel human cathepsin P is expressed at high levels in lung, liver and heart. Using antibodies raised against recombinant cathepsin P produced in Escherichia coli, a single protein band of 33 kDa was detected by immunoblotting an extract of human liver. By immunofluorescence, positive signals were observed in hepatocytes and Kupffer cells of liver, and the distal tubule cells of kidney showing mainly perimembranous distribution, indicating a role, as yet unknown, for this novel putative protease that is distinct from other cathepsins of the papain family.

Identification and molecular cloning of cathepsin P, a novel human putative cysteine protease of the papain family.

A cDNA encoding a novel human putative member of the papain family of cysteine peptidases has been cloned. The protease, named cathepsin P, is synthesized as a preproprotein. The presumed propeptide of 38 amino acids is followed by a 242-residue mature protein.

Tissue expression and immunolocalization of a novel human cathepsin P.

The mRNA of a novel human cathepsin P is expressed at high levels in lung, liver and heart. Using antibodies raised against recombinant cathepsin P produced in Escherichia coli, a single protein band of 33 kDa was detected by immunoblotting an extract of human liver. By immunofluorescence, positive signals were observed in hepatocytes and Kupffer cells of liver, and the distal tubule cells of kidney showing mainly perimembranous distribution, indicating a role, as yet unknown, for this novel putative protease that is distinct from other cathepsins of the papain family.

Identification and molecular cloning of cathepsin P, a novel human putative cysteine protease of the papain family.

A cDNA encoding a novel human putative member of the papain family of cysteine peptidases has been cloned. The protease, named cathepsin P, is synthesized as a preproprotein. The presumed propeptide of 38 amino acids is followed by a 242-residue mature protein.