Crystal structure of the platelet activator convulxin, a disulfide-linked alpha(4)beta(4) cyclic tetramer from the venom of Crotalus durissus terrificus

Convulxin (CVX), a C-type lectin, isolated from the venom of the South American rattlesnake Crotalus durissus terrificus, causes cardiovascular and respiratory disturbances and is a potent platelet activator which hinds to platelet glycoprotein GPVI. The structure of CVX has been solved at 2.4 Angstrom resolution to a crystallographic residual of 18.6% (R-free =26.4%). CVX is a disulfide linked heterodimer consisting of homologous alpha and beta chains. The heterodimers are additionally linked by disulfide bridges to form cyclic alpha(4)beta(4)heterotetramers. These domains exhibit significant homology to the carbohydrate-binding domains of C-type lectins, to the factor IX-binding protein (IX-bp), and to flavocetin-A (Fl-A) but sequence and Structural differences are observed in both the domains in the putative Ca2+ and carbohydrate binding regions. (C) 2003 Elsevier B.V. All rights reserved.

Chalcone-flavone tetramer and biflavones from Aristolochia ridicula

Biflavones and a chalcone flavone tetramer were isolated from the leaves of Aristolochia ridicula, together with proto-quercitol. Their structures were determined by spectroscopic methods. (c) 2005 Elsevier Ltd. All rights reserved.

Effective range from tetramer-dissociation data for cesium atoms

The shifts in the four-body recombination peaks, due to an effective range correction to the zero-range model close to the unitary limit, are obtained and used to extract the corresponding effective range of a given atomic system. The approach is applied to an ultracold gas of cesium atoms close to broad Feshbach resonances, where deviations of experimental values from universal model predictions are associated with effective range corrections. The effective range correction is extracted with a weighted average given by 3.9±0.8R vdW, where RvdW is the van der Waals length scale, which is consistent with the van der Waals potential tail for the Cs2 system. The method can be generally applied to other cold atom experimental setups to determine the contribution of the effective range to the tetramer dissociation position. © 2013 American Physical Society.

Range Corrections to Universal Tetramer Properties

The universal properties of weakly-bound tetramers close to the scaling limit are investigated by solving a subtracted set of Faddeev-Yakubovsky (FY) equations for identical bosons with a zero-range interaction. The solution demands a four-body scale independent of the trimer properties. Furthermore, the effect of a finite effective range is introduced in the FY equations, which we show produces results that are distinct from the scale variation. In particular range effects to two universal scaling functions for the tetramers are investigated. The correlation between successive tetramer energies corresponding to states within two Efimov trimer energies, proposed before and studied close to the unitary limit; and the correlation between the position of the four-atom recombination peaks. In this case, we found a shift in the scaling function due to the range, which can be associated to the shift of the data found for caesium atoms, with respect to zero-range calculations, due to a nonvanishing range in the actual experimental setups. © 2013 Springer-Verlag Wien.

Trimer-Tetramer Interwoven States in the Scaling Limit

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Identification of oligomers in polyethyleneterephthalate bottles for mineral water and fruit juice - Development and validation of a high-performance liquid chromatographic method for the determination of first series cyclic trimer

Cyclic oligomers were identified in PET bottles used for mineral water and fruit juice using MS and H-1 and C-13 NMR: a first series cyclic trimer, a first series cyclic tetramer, a first series cyclic dimmer and a second series cyclic trimer. An analytical method to determine first series cyclic trimer in these bottles was developed and validated, using HPLC. The first series cyclic trimer levels were 316-462 mg/100 g of PET bottle. (c) 2005 Elsevier B.V. All rights reserved.

Structure of chorismate synthase from Mycobacterium tuberculosis

In bacteria, fungi, plants, and apicomplexan parasites, the aromatics compounds, such as aromatics amino acids, are synthesized through seven enzymes from the shikimate pathway, which are absent in mammals. The absence of this pathway in mammals make them potential targets for development of new therapy against infectious diseases, such as tuberculosis, which is the world's second commonest cause of death from infectious disease. The last enzyme of shikimate pathway is the chorismate synthase (CS), which is responsible for conversion of the 5-enolpyruvylshikimate-3-phosphate to chorismate. Here, we report the crystallographic structure of CS from Mycobacterium tuberculosis (MtCS) at 2.65 angstrom resolution. The MtCS structure is similar to other CS structures, presenting beta-alpha-beta sandwich structural topology, in which each monomer of MtCS consists of a central helical core. The MtCS can be described as a tetramer formed by a dimer of dimers. However, analytical ultracentrifugation studies suggest the MtCS is a dimer with a more asymmetric shape than observed on the crystallographic dimer and the existence of a low equilibrium between dimer and tetramer. Our results suggest that the MtCS oligomerization is concentration dependent and some conformational changes must be involved on that event. (c) 2005 Elsevier B.V. All rights reserved.

Initial structural analysis of an alpha(4)beta(4) C-type lectin from the venom of Crotalus durissus terrificus

Convulxin, an alphabeta C-type lectin, is a potent platelet activator isolated from the venom of the South American rattlesnake Crotalus durissus terrificus. It is a 26.5 kDa alphabeta heterodimer consisting of two homologous disulfide-linked chains. The crystals belong to space group I4, with unit-cell parameters a = b = 131.61, c = 121.85 Angstrom, and diffraction data were collected to 2.7 Angstrom. The structure was solved by molecular replacement and the asymmetric unit contains two alphabeta heterodimers, each of which forms a disulfide-linked cyclic alpha(4)beta(4) tetramer in the unit cell. These alpha(4)beta(4) tetramers are stacked to form a large solvent channel.

Structural studies of prephenate dehydratase from Mycobacterium tuberculosis H37Rv by SAXS, ultracentrifugation, and computational analysis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Identification of a new quaternary association for legume lectins

Lotus tetragonolobus lectin (LTA) is a fucose-specific legume lectin. Although several studies report a diverse combination of biological activities for LTA, little is known about the mechanisms involved in L-fucosyl oligosaccharide recognition. The crystal structure of LTA at 2.0 angstrom resolution reveals a different legume lectin tetramer. Its structure consists of a homotetramer composed of two back-to-back GS4-like dimers arranged in a new mode, resulting in a novel tetramer. The LTA N-linked carbohydrate at Asn4 and the unusual LTA dimer-dimer interaction are related to its particular mode of tetramerization. In addition, we used small angle X-ray scattering to investigate the quaternary structure of LTA in solution and to compare it to the crystalline structure. Although the crystal structure of LTA has revealed a conserved metal-binding site, its L-fucose-binding site presents some punctual differences. Our investigation of the new tetramer of LTA and its fucose-binding site is essential for further studies related to cross-linking between LTA and complex divalent L-fucosyl carbohydrates. (C) 2007 Elsevier B.V. All rights reserved.

Crystallization, preliminary X-ray diffraction analysis and Patterson search of oxyhaemoglobin I from the wolf (Chrysocyon brachiurus)

Oxyhaemoglobin I isolated from the Brazilian wolf Chrysocyon brachiurus has been crystallized and X-ray diffraction data has been collected to 2.06 A resolution using a synchrotron-radiation source. Crystals were determined to belong to the space group P2(1)2(1)2(1) and preliminary structural analysis revealed the presence of one tetramer in the asymmetric unit. The structure was determined using standard molecular-replacement techniques and is currently being refined using maximum-likelihood protocols. This is the first haemoglobin isolated from a member of the Canidae family to be crystallized and it will provide further insights in the comparative biochemistry of vertebrate haemoglobins.

Rattlesnake hemoglobins: Functional properties and tetrameric stability

The present work analyzed the tetrameric stability of the hemoglobins from the rattlesnake C. durissus terrificus using analytical gel filtration chromatography, SAXS and osmotic stress. We show that the dissociation mechanism proposed for L. miliaris hemoglobin does not apply for these hemoglobins, which constitute stable tetramers even at low concentrations.

Four-boson scale near a Feshbach resonance

We show that an independent four-body momentum scale mu((4)) drives the tetramer binding energy for fixed trimer energy (or three-body scale mu((3))) and large scattering length (a). The three- and four-body forces from the one-channel reduction of the atomic interaction near a Feshbach resonance disentangle mu((4)) and mu((3)). The four-body independent scale is also manifested through a family of Tjon lines, with slope given by mu((4))/mu((3)) for a(-1) = 0. There is the possibility of a new renormalization group limit cycle due to the new scale.

Universality and Scaling Limit of Weakly-Bound Tetramers

The occurrence of a new limit cycle in few-body physics, expressing a universal scaling function relating the binding energies of two successive tetramer states, is revealed by considering a renormalized zero-range two-body interaction in bound state of four identical bosons. The tetramer energy spectrum is obtained by adding a boson to an Efimov bound state with energy B-3 in the unitary limit (for zero two-body binding energy or infinite two-body scattering length). Each excited N-th tetramer energy B-4((N)) is shown to slide along a scaling function as a short-range four-body scale is changed, emerging from the 3+1 threshold for a universal ratio B-4((N))/B-3 = 4.6, which does not depend on N. The new scale can also be revealed by a resonance in the atom-trimer recombination process.