Structure determination of the three disulfide bond isomers of alpha-conotoxin GI: A model for the role of disulfide bonds in structural stability

The three possible disulfide bonded isomers of alpha-conotoxin GI have been selectively synthesised and their structures determined by H-1 NMR spectroscopy. alpha-Conotoxin GI derives from the venom of Conus geographus and is a useful neuropharmacological tool as it selectively binds to the nicotinic acetylcholine receptor (nAChR), a ligand-gated ion channel involved in nerve signal transmission. The peptide has the sequence ECCNPACGRHYSC-NH2, and the three disulfide bonded isomers are referred to as GI(2-7;3-13), GI(2-13;3-7) and GI(2-3;7-13). The NMR structure for the native isomer GI(2-7;3-13) is of excellent quality, with a backbone pairwise RMSD of 0.16 Angstrom for a family of 35 structures, and comprises primarily a distorted 3(10),, helix between residues 5 to 11. The two non-native isomers exhibit multiple conformers in solution, with the major populated forms being different in structure both from each other and from the native form. Structure-activity relationships for the native GI(2-7;3-13) as well as the role of the disulfide bonds on folding and stability of the three isomers are examined. It is concluded that the disulfide bonds in alpha-conotoxin GI play a crucial part in determining both the structure and stability of the peptide. A trend for increased conformational heterogeneity was observed in the order of GI(2-7;3-13) < GI(2-13;3-7) < GI(2-3;7-13). It was found that the peptide bond joining Cys2 to Cys3 in GI(2-3;7-13) is predominantly trans, rather than cis as theoretically predicted. These structural data are used to interpret the varying nAChR binding of the non-native forms. A model for the binding of native GI(2-7;3-13) to the mammalian nAChR is proposed, with an alpha-subunit binding face made up of Cys2, Asn4, Pro5, Ala6 and Cys7 and a selectivity face, comprised of Arg9 and His10. These two faces orient the molecule between the alpha and delta subunits of the receptor. The structure of the CCNPAC sequence of the native GI(2-7;3-13) is compared to the structure of the identical sequence from the toxic domain of heat-stable enterotoxins, which forms part of the receptor binding region of the enterotoxins, but which has a different disulfide connectivity. (C) 1998 Academic Press Limited.

Power system sensitivity analysis for probabilistic small signal stability assessment in a deregulated environment

Deregulations and market practices in power industry have brought great challenges to the system planning area. In particular, they introduce a variety of uncertainties to system planning. New techniques are required to cope with such uncertainties. As a promising approach, probabilistic methods are attracting more and more attentions by system planners. In small signal stability analysis, generation control parameters play an important role in determining the stability margin. The objective of this paper is to investigate power system state matrix sensitivity characteristics with respect to system parameter uncertainties with analytical and numerical approaches and to identify those parameters have great impact on system eigenvalues, therefore, the system stability properties. Those identified parameter variations need to be investigated with priority. The results can be used to help Regional Transmission Organizations (RTOs) and Independent System Operators (ISOs) perform planning studies under the open access environment.

Eigenvalue sensitivity analysis for probabilistic small signal stability assessment

This paper presents a method to analyze the first order eigenvalue sensitivity with respect to the operating parameters of a power system. The method is based on explicitly expressing the system state matrix into sub-matrices. The eigenvalue sensitivity is calculated based on the explicitly formed system state matrix. The 4th order generator model and 4th order exciter system model are used to form the system state matrix. A case study using New England 10-machine 39-bus system is provided to demonstrate the effectiveness of the proposed method. This method can be applied into large scale power system eigenvalue sensitivity with respect to operating parameters.

Structure and function of plant toxins (with emphasis on cystine knot toxins)

Plant toxins are substances produced and secreted by plants to defend themselves against predators. In a broad sense, this includes all substances that have a toxic effect on targeted organisms, whether they are microbes, other plants, insects, or higher animals. Plant toxins have a diverse range of structures, from small organic molecules through to proteins. This review gives an overview of the various classes of plant toxins but focuses on an interesting class of protein-based plant toxins containing a cystine knot motif. This structural motif confers exceptional stability on proteins containing it and is associated with a wide range of biological activities. The biological activities and structural stability offer many potential applications in the pharmaceutical and agricultural fields. One particularly exciting prospect is in the use of protein-based plant toxins as molecular scaffolds for displaying pharmaceutically important bioactivities. Future applications of plant toxins are likely to involve genetic engineering techniques and molecular pharming approaches.

An Analytical Solution for the Elastic Lateral-Distortional Buckling of I-section Beams

Lateral-distortional buckling may occur in I-section beams with slender webs and stocky flanges. A computationally efficient method is presented in this paper to study this phenomenon. Previous studies on distortional buckling have been on the use of 3(rd) and 5(th) order polynomials to model the displacements. The present study provides an alternative way, using Fourier Series, to model the behaviour. Beams of different cross-sectional dimensions, load cases and restraint conditions are examined and compared. The accuracy and versatility of the method are verified by calibrating against the results of other published studies. The present method is believed to be a simple and efficient way of determining the buckling load and mode shapes of I-section beams that are susceptible to lateral-distortional buckling modes.

Distortional buckling of I-beams by finite element method

Distortional buckling, unlike the usual lateral-torsional buckling in which the cross-section remains rigid in its own plane, involves distortion of web in the cross-section. This type of buckling typically occurs in beams with slender web and stocky flanges. Most of the published studies assume the web to deform with a cubic shape function. As this assumption may limit the accuracy of the results, a fifth order polynomial is chosen here for the web displacements. The general line-type finite element model used here has two nodes and a maximum of twelve degrees of freedom per node. The model not only can predict the correct coupled mode but also is capable of handling the local buckling of the web.

Alpha-selenoconotoxins, a new class of potent alpha(7) neuronal nicotinic receptor antagonists

Disulfide bonds are important structural motifs that play an essential role in maintaining the conformational stability of many bioactive peptides. Of particular importance are the conotoxins, which selectively target a wide range of ion channels that are implicated in numerous disease states. Despite the enormous potential of conotoxins as therapeutics, their multiple disulfide bond frameworks are inherently unstable under reducing conditions. Reduction or scrambling by thiol-containing molecules such as glutathione or serum albumin in intracellular or extracellular environments such as blood plasma can decrease their effectiveness as drugs. To address this issue, we describe a new class of selenoconotoxins where cysteine residues are replaced by selenocysteine to form isosteric and non-reducible diselenide bonds. Three isoforms of alpha-conotoxin ImI were synthesized by t-butoxycarbonyl chemistry with systematic replacement of one([ Sec(2,8)] ImI or [Sec(3,12)] ImI), or both([Sec(2,3,8,12)] ImI) disulfide bonds with a diselenide bond. Each analogue demonstrated remarkable stability to reduction or scrambling under a range of chemical and biological reducing conditions. Three-dimensional structural characterization by NMR and CD spectroscopy indicates conformational preferences that are very similar to those of native ImI, suggesting fully isomorphic structures. Additionally, full bioactivity was retained at the alpha(7) nicotinic acetylcholine receptor, with each seleno-analogue exhibiting a dose-response curve that overlaps with wild-type ImI, thus further supporting an isomorphic structure. These results demonstrate that selenoconotoxins can be used as highly stable scaffolds for the design of new drugs.

Preparation and electrochemical properties of the layered material of LixVyO2 (x=0.86 and y=0.8)

Layered lithium-vanadium oxide with a composition of LixVyO2 (x = 0.86 and y = 0.8) was prepared by the hydrothermal reaction of V2O3 with LiOH center dot H2O at 180 degrees C. This material corresponds to a layered rhombohedral structure related to alpha-NaFeO2 in which the vanadium ions are disordered in alternate layers of octahedral 3a (0, 0, 0) and 3b (0, 0, 1/2) sites. The electrochemical properties of this Li0.86V0.8O2 material were investigated and compared with those of the layered Li0.96VO2 made by the conventional solid-state reaction. It was found that the electrochemical capacity and reversibility of the Li0.86V0.8O2 material are significantly improved compared to those of the Li0.96VO2 material; the reversible specific capacities of the Li/Li0.86V0.8O2 and Li/Li0.96VO2 systems are similar to 100 and similar to 50 mAh g(-1), respectively, under the current densities of 7.14 mA g(-1) over 20 charge-discharge cycles with a potential window of 1.50-4.50 V. Such a reversibility results from the structural stability of Li0.86V0.8O2, whereas the increase in the reversible specific capacity can be qualitatively interpreted in terms of the presence of vanadium vacancies in the structure. (c) 2005 The Electrochemical Society.

Effect of cultivation on soil C contents and saturated hydraulic conductivity

The irrigation of pasture with saline, Na-contaminated industrial wastewater typically results in an increase in soil ESP. From current knowledge (derived largely from cultivated agricultural soils), although these sodic soils are likely to remain stable whilst irrigated with effluent (due to the effluent’s large electrolyte concentration), during rainfall periods of low electrolyte concentration these soils would be expected to disperse. However, effluent irrigated pasture soils have been observed to maintain their structure even during intense rainfall events. Three soil types were collected (Sodosol, Vertosol and Dermosol), each with a cultivated/non-cultivated pair. The soils were equilibrated with various SAR solutions and then leached with deionised water to allow the measurement of saturated hydraulic conductivity (Ksat). At low SARs, Ksat tended to be greater in non-cultivated than cultivated soils and is attributable to a loss of structure associated with cultivation. In addition, as SAR increased, the reduction in relative Ksat tended to be significantly greater in cultivated than non-cultivated soils. The relatively rapid saturated hydraulic conductivity in the non-cultivated soils at large SARs is due to a greater aggregate stability due to greater soil C content. For the sustainable disposal of saline effluent, it is therefore necessary to ensure that soils remain undisturbed and preferably under pasture, thus maximising soil structural stability and hydraulic conductivity.

Bifurcation analysis of a dynamic power system with SVC devices

Bifurcation analysis is a very useful tool for power system stability assessment. In this paper, detailed investigation of power system bifurcation behaviour is presented. One and two parameter bifurcation analysis are conducted on a 3-bus power system. We also examined the impact of FACTS devices on power system stability through Hopf bifurcation analysis by taking static Var compensator (SVC) as an example. A simplified first-order model of the SVC device is included in the 3-bus sample system. Real and reactive powers are used as bifurcation parameter in the analysis to compare the system oscillatory properties with and without SVC. The simulation results indicate that the linearized system model with SVC enlarge the voltage stability boundary by moving Hopf bifurcation point to higher level of loading conditions. The installation of SVC increases the dynamic stability range of the system, however complicates the Hopf bifurcation behavior of the system

A study of the stability of subcycling algorithms in structural dynamics

Algorithms for explicit integration of structural dynamics problems with multiple time steps (subcycling) are investigated. Only one such algorithm, due to Smolinski and Sleith has proved to be stable in a classical sense. A simplified version of this algorithm that retains its stability is presented. However, as with the original version, it can be shown to sacrifice accuracy to achieve stability. Another algorithm in use is shown to be only statistically stable, in that a probability of stability can be assigned if appropriate time step limits are observed. This probability improves rapidly with the number of degrees of freedom in a finite element model. The stability problems are shown to be a property of the central difference method itself, which is modified to give the subcycling algorithm. A related problem is shown to arise when a constraint equation in time is introduced into a time-continuous space-time finite element model. (C) 1998 Elsevier Science S.A.

Structural analysis of three His32 mutants of DsbA: Support for an electrostatic role of His32 in DsbA stability

DsbA, a 21-kDa protein from Escherichia coli, is a potent oxidizing disulfide catalyst required for disulfide bond formation in secreted proteins. The active site of DsbA is similar to that of mammalian protein disulfide isomerases, and includes a reversible disulfide bond formed from cysteines separated by two residues (Cys3O-Pro31-His32-Cys33). Unlike most protein disulfides, the active-site disulfide of DsbA is highly reactive and the oxidized form of DsbA is much less stable than the reduced form at physiological pH. His32, one of the two residues between the active-site cysteines, is critical to the oxidizing power of DsbA and to the relative instability of the protein in the oxidized form. Mutation of this single residue to tyrosine, serine, or leucine results in a significant increase in stability (of similar to 5-7 kcal/mol) of the oxidized His32 variants relative to the oxidized wild-type protein. Despite the dramatic changes in stability, the structures of all three oxidized DsbA His32 Variants are very similar to the wild-type oxidized structure, including conservation of solvent atoms near the active-site residue, Cys3O. These results show that the His32 residue does not exert a conformational effect on the structure of DsbA. The destabilizing effect of His32 on oxidized DsbA is therefore most likely electrostatic in nature.