Movement Of Residues Research Articles

The 1.2 Å structure of a novel quorum-sensing protein, Bacillus subtilis LuxS

In bacteria, the regulation of gene expression in response to changes in cell density is called quorum sensing. The autoinducer-2 production protein LuxS, is involved in a novel quorum-sensing system and is thought to catalyse the degradation of S-ribosylhomocysteine to homocysteine and the autoinducer molecule 4,5-dihydroxy-2,3-pentadione. The crystal structure of Bacillus subtilis LuxS has been determined at 1.2 Å resolution, together with the binary complexes of LuxS with S-ribosylhomocysteine and homocysteine to 2.2 and 2.3 Å resolution, respectively. These structures show that LuxS is a homodimer with an apparently novel fold based on an eight-stranded β-barrel, flanked by six α-helices. Each active site contains a zinc ion coordinated by the conserved residues His54, His58 and Cys126, and includes residues from both subunits. S-ribosylhomocysteine binds in a deep pocket with the ribose moiety adjacent to the enzyme-bound zinc ion. Access to the active site appears to be restricted and possibly requires conformational changes in the protein involving the movement of residues 125–129 and those at the N terminus. The structure contains an oxidised cysteine residue in the active site whose role in the biological process of LuxS has not been determined. The autoinducer-2 signalling pathway has been linked to aspects of bacterial virulence and pathogenicity. The structural data on LuxS will provide opportunities for targeting this enzyme for the rational design of new antibiotics.

Reversibly bound and covalently attached ligands induce conformational changes in the omega loop, Cys69-Cys96, of mouse acetylcholinesterase.

We have used a combination of cysteine substitution mutagenesis and site-specific labeling to characterize the structural dynamics of mouse acetylcholinesterase (mAChE). Six cysteine-substituted sites of mAChE (Leu(76), Glu(81), Glu(84), Tyr(124), Ala(262), and His(287)) were labeled with the environmentally sensitive fluorophore, acrylodan, and the kinetics of substrate hydrolysis and inhibitor association were examined along with spectroscopic characteristics of the acrylodan-conjugated, cysteine-substituted enzymes. Residue 262, being well removed from the active center, appears unaffected by inhibitor binding. Following the binding of ligand, hypsochromic shifts in emission of acrylodan at residues 124 and 287, located near the perimeter of the gorge, reflect the exclusion of solvent and a hydrophobic environment created by the associated ligand. By contrast, the bathochromic shifts upon inhibitor binding seen for acrylodan conjugated to three omega loop (Omega loop) residues 76, 81, and 84 reveal that the acrylodan side chains at these positions are displaced from a hydrophobic environment and become exposed to solvent. The magnitude of fluorescence emission shift is largest at position 84 and smallest at position 76, indicating that a concerted movement of residues on the Omega loop accompanies gorge closure upon ligand binding. Acrylodan modification of substituted cysteine at position 84 reduces ligand binding and steady-state kinetic parameters between 1 and 2 orders of magnitude, but a similar substitution at position 81 only minimally alters the kinetics. Thus, combined kinetic and spectroscopic analyses provide strong evidence that conformational changes of the Omega loop accompany ligand binding.

Refined structures of β-ketoacyl-acyl carrier protein synthase III

β-Ketoacyl-acyl carrier protein synthase III (FabH) is a condensing enzyme that plays central roles in fatty acid biosynthesis. Three-dimensional structures of E. coli FabH in the presence and absence of ligands have been refined to 1.46 Å resolution. The structures of improved accuracy revealed detailed interactions involved in ligand binding. These structures also provided new insights into the FabH mechanism, e.g. the possible role of a water or hydroxyl anion in Cys112 deprotonation. A structure of the apo enzyme uncovered large conformational changes in the active site, exemplified by the disordering of four essential loops (84–86, 146–152, 185–217 and 305–307) and the movement of catalytic residues (Cys112 and His244). The disordering of the loops leads to greater than 50 % reduction in the FabH dimer interface, suggesting a dynamic nature for an unusually large portion of the dimer interface. The existence of a large solvent-accessible channel in the dimer interface as well as two cis-peptides ( cis-Pro88 and cis-Phe308) in two of the disordered loops may explain the observed structural instabilities.

Summary of well water sampling in California to detect pesticide residues resulting from nonpoint-source applications.

This report summarizes well sampling protocols, data collection procedures, and analytical results for the presence of pesticides in ground water developed by the California Department of Pesticide Regulation (DPR). Specific well sampling protocols were developed to meet regulatory mandates of the Pesticide Contamination Prevention Act (PCPA) of 1986 and to provide further understanding of the agronomic, chemical, and geographic factors that contribute to movement of residues to ground water. The well sampling data have formed the basis for the DPR's regulatory decisions. For example, a sampling protocol, the Four-Section Survey, was developed to determine if reported detections were caused by nonpoint-source agricultural applications, a determination that can initiate formal review and subsequent regulation of a pesticide. Selection of sampling sites, which are primarily rural domestic wells, was initially based on pesticide use and cropping patterns. Recently, soil and depth-to-ground water data have been added to identify areas where a higher frequency of detection is expected. In accordance with the PCPA, the DPR maintains a database for all pesticide well sampling in California with submission required by all state agencies and with invitations for submission extended to all local and federal agencies or other entities. To date, residues for 16 active ingredients and breakdown products have been detected in California ground water as a result of legal agricultural use. Regulations have been adopted for all detected parent active ingredients, and they have been developed regardless of the level of detection.

High Resolution Structure of the Phosphohistidine-activated Form of Escherichia coli Cofactor-dependent Phosphoglycerate Mutase

The active conformation of the dimeric cofactor-dependent phosphoglycerate mutase (dPGM) from Escherichia coli has been elucidated by crystallographic methods to a resolution of 1.25 A (R-factor 0.121; R-free 0.168). The active site residue His(10), central in the catalytic mechanism of dPGM, is present as a phosphohistidine with occupancy of 0.28. The structural changes on histidine phosphorylation highlight various features that are significant in the catalytic mechanism. The C-terminal 10-residue tail, which is not observed in previous dPGM structures, is well ordered and interacts with residues implicated in substrate binding; the displacement of a loop adjacent to the active histidine brings previously overlooked residues into positions where they may directly influence catalysis. E. coli dPGM, like the mammalian dPGMs, is a dimer, whereas previous structural work has concentrated on monomeric and tetrameric yeast forms. We can now analyze the sequence differences that cause this variation of quaternary structure.

Structural Changes Induced in p21 Ras upon GAP-334 Complexation as Probed by ESEEM Spectroscopy and Molecular-Dynamics Simulation

Background: The means by which the protein GAP accelerates GTP hydrolysis, and thereby downregulates growth signaling by p21 Ras, is of considerable interest, particularly inasmuch as p21 mutants are implicated in a number of human cancers. A GAP “arginine finger,” identified by X-ray crystallography, has been suggested as playing the principal role in the GTP hydrolysis. Mutagenesis studies, however, have shown that the arginine can only partially account for the 10 5-fold increase in the GAP-accelerated GTPase rate of p21. Results: We report electron spin-echo envelope modulation (ESEEM) studies of GAP-334 complexed with GMPPNP bound p21 in frozen solution, together with molecular-dynamics simulations. Our results indicate that, in solution, the association of GAP-334 with GTP bound p21 induces a conformational change near the metal ion active site of p21. This change significantly reduces the distances from the amide groups of p21 glycine residues 60 and 13 to the divalent metal ion. Conclusions: The movement of glycine residues 60 and 13 upon the binding of GAP-334 in solution provides a physical basis to interpret prior mutagenesis studies, which indicated that Gly-60 and Gly-13 of p21 play important roles in the GAP-dependent GTPase reaction. Gly-60 and Gly-13 may play direct catalytic roles and stabilize the attacking water molecule and β,γ-bridging oxygen, respectively, in p21. The amide proton of Gly-60 could also play an indirect role in catalysis by supplying a crucial hydrogen bonding interaction that stabilizes loop L4 and therefore the position of other important catalytic residues.

Identifying the adaptive mechanism in globular proteins: Fluctuations in densely packed regions manipulate flexible parts

A low-resolution structural model based on the packing geometry of α-carbons is utilized to establish a connection between the flexible and rigid parts of a folded protein. The former commonly recognizes a complementing molecule for making a complex, while the latter manipulates the necessary conformational change for binding. We attempt analytically to distinguish this control architecture that intrinsically exists in globular proteins. First with two-dimensional simple models, then for a native protein, bovine pancreatic trypsin inhibitor, we explicitly demonstrate that inserting fluctuations in tertiary contacts supported by the stable core, one can regulate the displacement of residues on loop regions. The positional fluctuations of the flexible regions are annihilated by the rest of the protein in conformity with the Le Chatelier–Braun principle. The results indicate that the distortion of the principal nonbonded contacts between highly packed residues is accompanied by that of the slavery fluctuations that are widely distributed over the native structure. These positional arrangements do not appear in a reciprocal relation between a perturbation and the associated response; the effect of a movement of residue i on residue j is not equal to that of the same movement of residue j on residue i.

Steady state and picosecond time-resolved fluorescence studies on native, desulpho and deflavo xanthine oxidase

Steady state and time-resolved fluorescence studies on native, desulpho and deflavo xanthine oxidase (XO) have been carried out to investigate the conformational changes associated with the replacement of the molybdenum double bonded sulphur by oxygen and the removal of the flavin adenine dinucleotide (FAD). The steady state quenching experiments of the intrinsic tryptophan residues of the enzyme show that all the nine tryptophans are accessible to neutral quencher, acrylamide, in the native as well as desulpho and deflavo enzymes. However, the number of the tryptophan residues accessible to the ionic quenchers, potassium iodide and cesium chloride, increases upon removal of the FAD centre from the enzyme. This indicates that two tryptophan residues move out from the core of the enzyme to the solvent upon the removal of the FAD. The time-resolved fluorescence studies were carried out on the native, desulpho and deflavo XO by means of the time-correlated single photon counting technique, and the data were analysed by discrete exponential and maximum entropy methods. The results show that the fluorescence decay curve fitted best to a three-exponential model with lifetimes τ 1=0.4, τ 2=1.4 and τ 3=3.0 ns for the native and desulpho XO, and τ 1=0.7, τ 2=1.7 and τ 3=4.8 ns for the deflavo XO. The replacement of the molybdenum double bonded sulphur by oxygen in the desulpho enzyme does not cause any significant change of the lifetime components. However, removal of the FAD centre causes a significant change in the shortest and longest lifetime components indicating a conformational change in the deflavo XO possibly in the flavin domain. Decay-associated emission spectra at various emission wavelengths have been used to determine the origin of the lifetimes. The results show that τ 1 and τ 3 of the native and desulpho XO originate from the tryptophan residues which are completely or partially accessible to the solvent but τ 2 corresponds to those residues which are buried in the core of the enzyme and not exposed to the solvent. For deflavo enzyme, τ 2 is red shifted compared to the native enzyme indicating the movement of tryptophan residues from the core of the enzyme to the solvents.

A preliminary examination of the translocation of microencapsulated cyfluthrin following applications to the perimeter of residential dwellings

Methods have been developed to monitor the translocation of microencapsulated cyfluthrin following perimeter applications to residential dwellings. A pilot study was implemented to determine both the potential for application spray to drift away from dwellings and the intrusion of residues into homes following perimeter treatments. Residential monitoring included measuring spray drift using cellulose filter paper and the collection of soil samples from within the spray zone. In addition, interior air was monitored using fiberglass filter paper as a sorbent medium and cotton ball swabs were used to collect surface wipes. Fortification of matrixes resulted in recoveries of >90%. Spray drift was highest at the point of application and declined to low but measurable levels 9.1m from the foundations of dwellings. Soil residues declined to low, but measurable levels by 45 days post‐application. No cyfluthrin was measured from indoor air; however, some interior surfaces had detectable levels of cyfluthrin until three days post‐application. Findings indicate that spray drift resulting from perimeter applications might contaminate non‐target surfaces outside the spray zone. Soil borne residues may serve as persistent sources for human exposure and potentially intrude into dwellings through the activities of occupants and pets. Residues do not appreciably translocate through air and consequently inhalation is not a likely route for human exposure. Surface residues detected indoors suggest that the physical movement of residues from the exterior to the interior might be a viable route of movement of residues following this type of application.

Local Movement in the S2 Region of the Voltage-Gated Potassium Channel hKv2.1 Studied Using Cysteine Mutagenesis

The positively charged S4 region of voltage-dependent potassium channels moves outward during depolarization, leading to channel opening, but possible movement of the negatively charged S2 region may be more complex. Here we have studied possible movement of the S2 region of the slowly activating human voltage-dependent potassium channel hKv2.1. For this, cysteine mutants in the S2 region were expressed in Xenopus oocytes by injection of cRNA. Whole-cell currents were measured using the two-electrode voltage-clamp technique, and the effect of the membrane-impermeable cysteine-binding reagent parachloromercuribenzenesulfonate (PCMBS) was studied. For mutant S223C (located just outside the membrane in the S2 region), PCMBS inhibited currents and caused faster deactivation of tail currents. The time course of reactivity of PCMBS on tail current amplitudes was faster at more negative holding potentials. There was no effect of PCMBS on potassium channel currents for mutants D225C, N226C, A230C, and V232C. These data suggest that residue S223 is exposed to the extracellular phase at normal resting potentials, making it accessible to PCMBS, but upon depolarization there is a conformational change, making it less accessible, possibly by a local rather than global movement of S2 residues into the membrane. Voltage-dependent movements of nearby residues could also explain the results.

A new twist in the saga of charge movement in voltage-dependent ion channels.

Despite the many discrepancies in results and the different experimental methods used, both papers arrive at the same conclusion. S4 segments rotate when depolarized, and this rotation moves charged residues from an intracellular vestibule to an extracellular vestibule (Figure 2BFigure 2B). One reason this mechanism is so appealing is that the residues most important for charge transfer, the outermost four charged residues, lie along one face of an α helix. Note that in order for this mechanism to account for charge transfer, the direction of the electric field cannot be oriented parallel to the plane of the membrane, as implied for example by the shaded regions in Figure 1CFigure 1C, but more perpendicular to the membrane (Figure 2BFigure 2B). This reorientation of the electric field is a consequence of the topology of the aqueous vestibules surrounding the S4 segment.One prediction of these models is that the residues on the backside of this stripe of positive charge, i.e., the neutral residues between the charged residues in the primary sequence, should move in the opposite direction, from extracellular to intracellular during a depolarization. This does not occur. The residues between the second and third arginines in the primary sequence are in an intracellular compartment at hyperpolarized voltages (Figure 1CFigure 1C). This does not eliminate the possibility that S4 rotates. It means that the gating pore also undergoes conformational changes as the S4 segment moves. This fact is also apparent from accessibility studies, which suggest that the length of the gating pore changes with depolarization (Figure 1CFigure 1C).What message can we take home about S4 movement? As attractive as it is, the claim for a rotation as large as 180° is weak. Furthermore, even if a smaller rotation contributes to charge movement, it is likely to be only one of the movements of S4 and the protein surrounding it. Perhaps the most important conclusion supported by both studies is that the movements of the S4 segment are very small, as one might expect for a conformational change that has to be fast enough to underlie the gating currents.The raison d'etre of an S4 segment is not charge movement; it is the control of the activation gates at the bottom of the S6 segments. How are these movements coupled? One possibility is that movement of the S4 segment tugs or twists the linkers that connect it to S3 and S5 segments, and these segments transmit this movement to the S6 segment. Another possibility is that S4 and S6 are like entwined lovers. One cannot move without a compensatory rearrangement of the other. For example, S4 rotation may cause S6 rotation, a movement that underlies the gating of a bacterial potassium channel (Perozo et al. 1999xPerozo, E., Cortes, D.M., and Cuello, L.G. Science. 1999; 285: 73–78Crossref | PubMed | Scopus (461)See all ReferencesPerozo et al. 1999). The answers to these mysteries will surely unfold with the next episode of conformational studies of voltage-dependent gating.*E-mail: richard.horn@mail.tju.edu.

Metais pesados em solos de área de rejeitos de indústria de processamento de zinco

Apesar de a poluição do solo ser amplamente constatada, estudos sobre as concentrações e formas dos metais pesados em solos e seus efeitos no ecossistema são ainda pouco enfatizados, principalmente em condições tropicais. O presente estudo foi desenvolvido em área de rejeitos de uma unidade de extração e industrialização de zinco pertencente à Companhia Mineira de Metais-CMM, em Três Marias (MG), com o objetivo de avaliar as quantidades e formas de metais pesados em sete locais representativos da área. Os locais foram selecionados para amostragem, baseando-se em diferenças na forma de contaminação, aspectos de solo, topografia e vegetação. Realizaram-se análises químicas de fracionamento de metais pesados e extrações simples com DTPA e Mehlich-1, buscando determinar os teores e formas dos metais na superfície e em profundidade e fazer inferências sobre o potencial de risco ambiental desses metais. Os teores totais dos metais nas camadas superficiais dos locais estudados foram, em média, de: 13.533 mg kg-1, para Zn; 170 mg kg-1, para Cd; 865 mg kg-1, para Cu, e 612 mg kg-1, para Pb, enquanto os teores trocáveis (MgCl2) nessas mesmas camadas variaram de 231 a 1.407 mg kg-1, para Zn; 14 a 390 mg kg-1, para Cd, e 11 a 33 mg kg-1, para Pb; o Cu raramente ocorreu nesta forma. Os solos dos locais estudados, com teores excessivos de Zn, Cd, Cu e Pb, são considerados poluídos. Em determinados locais contaminados por escoamento (superficial e subsuperficial) e arraste de material de solo e rejeito, a poluição mostrou-se mais evidente nas camadas superficiais. No local da ustulação, verificou-se maior percentagem de Zn trocável em todos os níveis de profundidade. Nos demais locais, o Cd apresentou também grandes concentrações nas formas trocáveis, razão por que tal elemento oferece maior risco de contaminação ambiental. De modo geral, a ocorrência de Zn foi maior nas formas carbonato e residual, enquanto o Cd predominou nas formas trocáveis e o Cu e Pb nas formas residuais. A extração dos metais por Mehlich-1 e DTPA correlacionou-se significativamente com a soma das extrações com MgCl2 e NaOAc do fracionamento, para praticamente todos os elementos analisados. Por essa razão, recomendam-se futuros estudos com aqueles extratores em termos de mensuração de metais potencialmente disponíveis ao ambiente.

Manganese Is Essential for Catalytic Activity ofEscherichia coliAgmatinase

PurifiedEscherichia coliagmatinase (EC 3.5.3.11) expressed the same activity in the absence or presence of added Mn2+(0-5mM). However, it was strongly inhibited by Co2+, Ni2+, and Zn2+and almost half inactivated by EDTA. Partial inactivation by EDTA yielded enzyme species containing 0.85 ± 0.1 Mn2+/subunit, and it was accompanied by a decrease in intensity of fluorescence emission and a red shift from the emission maximum of 340 nm to 346 nm, indicating the movement of tryptophane residues to a more polar environment. The activity and fluorescence properties of fully activated agmatinase were restored by incubation of dialysed species with Mn2+. Manganese-free species, obtained by treatment with EDTA and guanidinium chloride (3 M), were active only in the presence of added Mn2+. Results obtained, which represent the first demonstration of the essentiality of Mn2+for agmatinase activity, are discussed in connection with a possible binuclear metal center in the enzyme.

The crystal structure of a novel bacterial adenylyltransferase reveals half of sites reactivity.

Phosphopantetheine adenylyltransferase (PPAT) is an essential enzyme in bacteria that catalyses a rate-limiting step in coenzyme A (CoA) biosynthesis, by transferring an adenylyl group from ATP to 4'-phosphopantetheine, yielding dephospho-CoA (dPCoA). Each phosphopantetheine adenylyltransferase (PPAT) subunit displays a dinucleotide-binding fold that is structurally similar to that in class I aminoacyl-tRNA synthetases. Superposition of bound adenylyl moieties from dPCoA in PPAT and ATP in aminoacyl-tRNA synthetases suggests nucleophilic attack by the 4'-phosphopantetheine on the alpha-phosphate of ATP. The proposed catalytic mechanism implicates transition state stabilization by PPAT without involving functional groups of the enzyme in a chemical sense in the reaction. The crystal structure of the enzyme from Escherichia coli in complex with dPCoA shows that binding at one site causes a vice-like movement of active site residues lining the active site surface. The mode of enzyme product formation is highly concerted, with only one trimer of the PPAT hexamer showing evidence of dPCoA binding. The homologous active site attachment of ATP and the structural distribution of predicted sequence-binding motifs in PPAT classify the enzyme as belonging to the nucleotidyltransferase superfamily.

Molecular dynamics study of femtosecond events in photoactive yellow protein after photoexcitation of the chromophore.

Molecular dynamics simulations were carried out to study what happens in a photoreceptor protein, photoactive yellow protein (PYP), immediately after the vertical transition of the chromophore from the ground to the excited state. A photon absorption simulation was performed to investigate the movement of amino acid residues upon photoexcitation. To calculate the excited state of the chromophore, SCF-CI calculation was carried out with INDO/S Hamiltonian. We observed that some amino acid residues have strong interactions with the chromophore. Most of these amino acid residues are conserved in PYPs from three different species of bacteria. This observation indicates the biological importance of these residues.

Soil Organic Matter Dynamics after Addition of Nitrogen‐15‐Labeled Leucaena and Dactyladenia Residues

AbstractQuantification of the movement of residue N through various biologically meaningful soil organic matter (SOM) fractions is essential in low‐input tropical cropping systems for the development of management practices aimed at optimizing N‐use efficiency. The flow of N derived from 15N‐labeled leucaena [Leucaena leucocephala (Lam.) de Wit] and dactyladenia [Dactyladenia barteri (Hook f ex Oliv.) Engl.] leaf residues was followed through the SOM of different particle‐size fractions during 858 d. A bioassay study with maize (Zea mays L.) was carried out to assess the availability of residue‐derived N (RDN) incorporated in the particle‐size fractions. At 53 d after residue application (DAA), most of the RDN was found in the fraction between 0.250 and 2 mm (O250) in the leucaena treatment and in the fraction >2 mm and the surface litter (O2000 + SL) in the dactyladenia treatment. The RDN content of all fractions <0.250 mm was significantly higher in the leucaena than in the dactyladenia treatment up to 471 DAA. The proportion of the total amount of RDN recovered in the fractions <2 mm shifted toward the smaller particle‐size classes near the end of the experiment. Highly significant (P < 0.001) relationships between RDN present in the particulate organic matter (POM) and uptake of RDN by maize indicated the relatively high availability of RDN in the POM. Fractionating the SOM pool into different particle‐size classes yielded useful information on the relative contributions of the different SOM fractions to N turnover and availability. This information is needed in identifying SOM pools with direct relevance to plant nutrition in low‐input cropping systems.

Molecular Dynamics Simulation of the Excited‐State Dynamics of Bacteriorhodopsin

Abstract— The excited‐state dynamics of bacteriorhodopsin was studied by molecular dynamics simulation. For the purpose of suppressing large displacement of amino acid residues on the surface of bacteriorhodopsin, positional restraints were imposed on these residues. A new method was developed to investigate the movement of amino acid residues upon photoexcitation and their role on the ultrafast photoisomerization of the chromophore. The structural change of bacteriorhodopsin was then traced up to 200 fs, i.e. until the formation of the intermediate I. We found that when all the conjugated bonds of the chromophore were allowed to twist freely in the excited state, many bonds including the C13=C14 bond twist in large scale within 100 fs. When only the C13=C14 bond and the single bonds were allowed to twist freely, the twisting took place at most 20° within 200 fs. From these results, it is claimed that a special potential surface is provided for the C13=C14 bond twisting by the protein environment in the course of the isomerization reaction, giving rise to the specific, ultrafast photoisomerization of bacteriorhodopsin. As a trace of such a mechanism, we observed that several functionally important residues incuding Asp85, Asp212 and Tyr185 responded quickly to the photoexcitation of the chromophore.

An analytical model for the assessment of pesticide exposure levels in soils and groundwater

The movement and degradation of pesticide residues in soils and groundwater are complex processes affected by soil physical, (bio)chemical, and hydrogeological properties, climatic conditions, and agricultural practices. This work presents a physically-based analytical model suitable for long-term predictions of pesticide concentrations in groundwater. The primary interest is to investigate the impact of soil environment, related physical and (bio)chemical processes, especially, volatilization, crop uptake, and agricultural practices on long-term vulnerability of groundwater to contamination by pesticides. The soil is separated into root and intermediate vadose zones, each with uniform properties. Transport in each soil zone is modeled on the basis of complete mixing, by spatial averaging the related point multiphase-transport partial differential equation (i.e., linear-reservoir models). Transport in the aquifer, however, is modeled by a two-dimensional advection-dispersion transport equation, considering adsorption and first-order decay rate. Vaporization in the soil is accounted for by assuming liquid-vapor phase partitioning using Henry's law, and vapor flux (volatilization) from the soil surface is modeled by diffusion through an air boundary layer. Sorption of liquid-phase solutes by crops is described by a linear relationship which is valid for first-order (passive) crop uptake. The model is applied to five pesticides (atrazine, bromacil, chlordane, heptachlor, and lindane), and the potential for pesticide contamination of groundwater is investigated for sandy and clayey soils. Simulation results show that groundwater contamination can be substantially reduced for clayey soil environments, where bio(chemical) degradation and volatilization are most efficient as natural loss pathways for the pesticides. Also, uptake by cross can be a significant mechanism for attenuating exposure levels in ground-water especially in a sandy soil environment, and for relatively persisting pesticides. Further, simulations indicate that changing agricultural practices can have a profound effect on vulnerability of groundwater to mobile and relatively persisting pesticides.

Voltage dependence of mouse acetylcholine receptor gating: different charge movements in di-, mono- and unliganded receptors.

1. The voltage dependence of binding and gating in wild-type and mutant recombinant mouse nicotinic acetylcholine receptors (AChRs) was examined at the single-channel level. 2. The closing rate constant of diliganded receptors decreased e-fold with approximately 66 mV hyperpolarization in both wild-type (adult and embryonic) and mutant receptors. The opening rate constant of a mutant receptor (alpha Y93F) was not voltage dependent. 3. The voltage dependence of closing in monoliganded receptors was examined in several receptors having a mutation in the binding site (alpha G153S) or pore region (alpha L251C and epsilon T264P). The closing rate constant of these monoliganded receptors decreased e-fold with approximately 124 mV hyperpolarization. 4. The voltage dependence of closing and opening in unliganded receptors was examined in two receptors having a mutation in the pore region (alpha L251C and epsilon T264P). Neither the closing nor the opening rate constants of unliganded receptors were voltage dependent. 5. If z if the amount of charge that moves during channel closure and delta is the distance (as a fraction of the electric field) that the charge moves, we conclude that z delta = 0.4 in diliganded receptors, 0.2 in monoliganded receptors, and 0.0 in unliganded receptors. It is likely that charges on the protein, rather than the agonist molecule, move z delta = 0.2 after each ACh molecule has bound. 6. The results suggest that unliganded openings arise from a local, concerted change in the structure of the pore (channel opening) that does not involve the net movement of charged residues. We speculate that as a consequence of agonist binding, charged moieties in the protein change their disposition so that they move with respect to the electric field when the channel gates. The results are consistent with the idea that there is semi-independent movement of distinct domains during AChR gating.

Evaluation of three pesticide leaching models with experimental data for alachlor, atrazine and metribuzin

SummaryThe persistence and movement of residues of alachlor, alrazine and metribuzin were measured in a mini‐lysimeter system in the field. This comprised a number of soil columns (11 cm diametert; 30 cm long), and permitted the vertical distribution of residues to be determined at. intervals alter application and the collection and analysis of leaehale water. Laboratory experiments were also performed to determine the degradation rates of the three herbicides and their strengths of adsorption by the test soil. The results showed an order of degradation rate of metribuzin> alachlor>atrazine and an order of adsorption of alacblor>atrazine>melribuzin. Movement of residues in the soil columns and concentrations in the leachate were inversely related to the strength of adsorption. Parameters derived from the laboratory data were used in conjunction with weather data for the period of the field experiment in three mathematical models of pesticide leaching: VARLEACH, LEACHP and PRZM2. In most instances, the models gave acceptable predictions of the distribution of residues in soil. This was particularly so for the less mobile compound alachlor. With the most mobile compound, metribuzin, residues were not well predicted at the later sampling dates. All three models gave accurate predictions of the volumes of drainage water, but none of them predicted the concentrations of herbicide in the leachate, presumably because they do not take account of preferential flow pathways of water and solute in the soil.