scholarly journals Spatial positioning of CFTR’s pore-lining residues affirms an asymmetrical contribution of transmembrane segments to the anion permeation pathway

2016 ◽  
Vol 147 (5) ◽  
pp. 407-422 ◽  
Author(s):  
Xiaolong Gao ◽  
Tzyh-Chang Hwang
Keyword(s):  
Metal Ion ◽  
Computational Models ◽  
Spatial Relationship ◽  
Chelating Agent ◽  
Inhibitory Effect ◽  
Physical Constraints ◽  
Transmembrane Segments ◽  
Narrow Region ◽  
Anion Permeation ◽  
Pore Lining

The structural composition of CFTR’s anion permeation pathway has been proposed to consist of a short narrow region, flanked by two wide inner and outer vestibules, based on systematic cysteine scanning studies using thiol-reactive probes of various sizes. Although these studies identified several of the transmembrane segments (TMs) as pore lining, the exact spatial relationship between pore-lining elements remains under debate. Here, we introduce cysteine pairs in several key pore-lining positions in TM1, 6, and 12 and use Cd2+ as a probe to gauge the spatial relationship of these residues within the pore. We find that inhibition of single cysteine CFTR mutants, such as 102C in TM1 or 341C in TM6, by intracellular Cd2+ is readily reversible upon removal of the metal ion. However, the inhibitory effect of Cd2+ on the double mutant 102C/341C requires the chelating agent dithiothreitol (DTT) for rapid reversal, indicating that 102C and 341C are close enough to the internal edge of the narrow region to coordinate one Cd2+ ion between them. We observe similar effects of extracellular Cd2+ on TM1/TM6 cysteine pairs 106C/337C, 107C/337C, and 107C/338C, corroborating the idea that these paired residues are physically close to each other at the external edge of the narrow region. Although these data paint a picture of relatively symmetrical contributions to CFTR’s pore by TM1 and TM6, introducing cysteine pairs between TM6 and TM12 (348C/1141C, 348C/1144C, and 348C/1145C) or between TM1 and TM12 (95C/1141C) yields results that contest the long-held principle of twofold pseudo-symmetry in the assembly of ABC transporters’ TMs. Collectively, these findings not only advance our current understanding of the architecture of CFTR’s pore, but could serve as a guide for refining computational models of CFTR by imposing physical constraints among pore-lining residues.

Sorption Profile and Thermodynamic Characteristics of Nitrosonaphthol Functionalized Sorbent for Metal Ion Enrichment

2005 ◽  
Vol 70 (9) ◽  
pp. 1341-1356 ◽  
Author(s):  
Saima Q. Memon ◽  
Muhammad I. Bhanger ◽  
Muhammad Y. Khuhawar
Keyword(s):  
Metal Ion ◽  
Tap Water ◽  
Thermodynamic Characteristics ◽  
Chelating Agent ◽  
Standard Addition ◽  
Absorption Spectrometry ◽  
Sorption Data ◽  
Maximum Sorption ◽  
Styrene Divinylbenzene ◽  
Two Phases

A simple and reliable method has been developed using styrene-divinylbenzene-based polymeric material containing 1-nitroso-2-naphthol as chelating agent, to concentrate ultratrace amounts of Ni(II) and Cu(II) ions in aqueous samples. Sorption of both the ions on the new synthetic resin under static and dynamic conditions has been investigated. The sorption has been optimized with respect to pH, shaking and contact time of two phases. Maximum sorption has been achieved from solution of pH 5-8 after 8 min of agitation. Total saturation capacities were 516 ± 2 and 316 ± 2.5 μmol g-1 for Ni(II) and Cu(II) ions, respectively. The lowest concentration for quantitative recovery (98 ± 1%) is 1.33 and 5 ppb with the preconcentration factor of 750 and 200 for Ni(II) and Cu(II), respectively. Monitoring of the influence of diverse ions on the sorption of metal ions has revealed that phosphate, hydrogencarbonate and citrate reduce the sorption to some extent. Under optimum conditions the sorption data followed Langmuir, Freundlich, and Dubinin-Radushkevich isotherms. The kinetics and thermodynamics of sorption are studied in detail. The sorption procedure is utilized to preconcentrate these ions prior to their determination in tea, human hair, and tap water samples by atomic absorption spectrometry using direct and standard addition methods.

Potentiation by L-cysteine of the bactericidal effect of hydrogen peroxide in Escherichia coli

1982 ◽  
Vol 152 (1) ◽  
pp. 81-88
Author(s):  
E H Berglin ◽  
M B Edlund ◽  
G K Nyberg ◽  
J Carlsson
Keyword(s):  
Escherichia Coli ◽  
Hydrogen Peroxide ◽  
Metal Ion ◽  
Chelating Agent ◽  
Fold Increase ◽  
Bactericidal Effect ◽  
Anaerobic Conditions ◽  
Dna Breakage ◽  
E Coli ◽  
K 12

Under anaerobic conditions an exponentially growing culture of Escherichia coli K-12 was exposed to hydrogen peroxide in the presence of various compounds. Hydrogen peroxide (0.1 mM) together with 0.1 mM L-cysteine or L-cystine killed the organisms more rapidly than 10 mM hydrogen peroxide alone. The exposure of E. coli to hydrogen peroxide in the presence of L-cysteine inhibited some of the catalase. This inhibition, however, could not fully explain the 100-fold increase in hydrogen peroxide sensitivity of the organism in the presence of L-cysteine. Of other compounds tested only some thiols potentiated the bactericidal effect of hydrogen peroxide. These thiols were effective, however, only at concentrations significantly higher than 0.1 mM. The effect of L-cysteine and L-cystine could be annihilated by the metal ion chelating agent 2,2'-bipyridyl. DNA breakage in E. coli K-12 was demonstrated under conditions where the organisms were killed by hydrogen peroxide.

scholarly journals Allosteric potentiation of a ligand-gated ion channel is mediated by access to a deep membrane-facing cavity

2018 ◽  
Vol 115 (42) ◽  
pp. 10672-10677 ◽  
Author(s):  
Stephanie A. Heusser ◽  
Marie Lycksell ◽  
Xueqing Wang ◽  
Sarah E. McComas ◽  
Rebecca J. Howard ◽  
...  
Keyword(s):  
Binding Site ◽  
Binding Sites ◽  
Inhibitory Effect ◽  
Allosteric Modulation ◽  
Bacterial Protein ◽  
Detailed Model ◽  
Differential Motion ◽  
State Dependent ◽  
The Common ◽  
Pore Lining

Theories of general anesthesia have shifted in focus from bulk lipid effects to specific interactions with membrane proteins. Target receptors include several subtypes of pentameric ligand-gated ion channels; however, structures of physiologically relevant proteins in this family have yet to define anesthetic binding at high resolution. Recent cocrystal structures of the bacterial protein GLIC provide snapshots of state-dependent binding sites for the common surgical agent propofol (PFL), offering a detailed model system for anesthetic modulation. Here, we combine molecular dynamics and oocyte electrophysiology to reveal differential motion and modulation upon modification of a transmembrane binding site within each GLIC subunit. WT channels exhibited net inhibition by PFL, and a contraction of the cavity away from the pore-lining M2 helix in the absence of drug. Conversely, in GLIC variants exhibiting net PFL potentiation, the cavity was persistently expanded and proximal to M2. Mutations designed to favor this deepened site enabled sensitivity even to subclinical concentrations of PFL, and a uniquely prolonged mode of potentiation evident up to ∼30 min after washout. Dependence of these prolonged effects on exposure time implicated the membrane as a reservoir for a lipid-accessible binding site. However, at the highest measured concentrations, potentiation appeared to be masked by an acute inhibitory effect, consistent with the presence of a discrete, water-accessible site of inhibition. These results support a multisite model of transmembrane allosteric modulation, including a possible link between lipid- and receptor-based theories that could inform the development of new anesthetics.

scholarly journals Animacy and the prediction of behaviour

2020 ◽  
Author(s):  
Johannes Schultz ◽  
Chris D. Frith
Keyword(s):  
Visual Cues ◽  
Computational Models ◽  
Action Observation ◽  
Critical Role ◽  
Mirror Neuron ◽  
Neuron Network ◽  
Short Term ◽  
Prior Beliefs ◽  
Physical Constraints ◽  
Intentional Agents

To survive, all animals need to predict what other agents are going to do next. The first step is to detect that an object is an agent and, if so, how sophisticated it is. To this end, visual cues are especially important: the form of the agent and the nature of its movements. Once identified, the movements of an agent, however sophisticated, can be anticipated in the short term on the basis of purely physical constraints, but, in the longer term, it is useful to take account of the agent’s goals and intentions. Goal directed agents are marked by the rationality of their movements, reaching their goals by the shortest or least effortful path. Observing goal directed behaviour activates the brain’s action observation/mirror neuron network. The observer’s own action generating mechanism has an important role in predicting future movements of goal directed agents.Intentions have a critical role in determining actions when agents interact with other agents. In such interactions, movements can become communicative rather than directed to immediate goals. Also, each agent can be trying to predict the behaviour of the other, leading to a recursive arms race. It is difficult to infer intentional behaviour from movement kinematics and interpretation is much more dependent upon prior beliefs about the agent. When people believe that they are interacting with an intentional agent, the brain’s mentalising system is activated as the person tries to assess the degree of sophistication of the agent. Several biologically-constrained computational models of action recognition are available, but equivalent models for understanding intentional agents remain to be developed.

THE EFFECT OF BETA-DECAY ON THE EXCHANGE PROPERTIES OF THE RARE EARTH–EDTA COMPLEX IONS

1961 ◽  
Vol 39 (5) ◽  
pp. 1049-1053 ◽  
Author(s):  
P. Glentworth ◽  
R. H. Betts
Keyword(s):  
Rare Earth ◽  
Metal Ion ◽  
Chemical Reactivity ◽  
Sudden Change ◽  
Chelating Agent ◽  
Beta Particle ◽  
Central Metal ◽  
Complex Ions ◽  
Rare Earth Ion ◽  
The Rare Earth

It is shown that the rare earth ion Yb3+ is very resistant towards ordinary thermal exchange when it is complexed with the chelating agent EDTA in aqueous solution. However, when the complexed rare earth atom, as the 1.8-h Yb-177, emits a beta-particle, the daughter atom Lu-177 escapes readily from the chelate structure. Nuclear recoil arising from the beta-particle emission is shown not to be the cause of the escape of the daughter atom. It is suggested that the observed lability of the daughter atom is a result of a high degree of chemical reactivity of the chelate ion arising from the sudden change in atomic number of the central metal ion of the chelate structure.

scholarly journals Determination of Traces of Pd(II) in Spiked Samples by Using 3,4-Dihydroxybenzaldehydeisonicotinol-hydrazone as a Chelating Agent with UV Visible Spectrophotometer

2011 ◽  
Vol 8 (1) ◽  
pp. 217-225 ◽  
Author(s):  
S. Lakshmi Narayana ◽  
C. Ramachandraiah ◽  
A. Varada Reddy ◽  
Dongyeun Lee ◽  
Jaesool Shim
Keyword(s):  
Metal Ion ◽  
Regression Coefficient ◽  
Chelating Agent ◽  
Molar Absorptivity ◽  
Inexpensive Method ◽  
Colored Complex ◽  
Major Cations ◽  
Uv Visible ◽  
Metal Ligand

A simple, rapid, sensitive and inexpensive method has been developed for the determination of trace amounts of palladium(II) using 3,4-dihydroxybenzaldehydeisonicotinoylhydrazone (3,4-DHBINH). The metal ion gives a yellow colored complex with 3,4-DHBINH in acetate buffer of pH 3.0 with 1:1 (metal: ligand) composition. The complex shows maximum absorption at 380 nm. Beer’s law is obeyed in the range 0.5-20.0 ppm of Pd(II). The molar absorptivity, Sandell’s sensitivity and detection limit were found to be 0.53×104L mol-1cm-1, 0.02 μg cm-2and 0.0948 μg mL-1, respectively. The correlation coefficient and regression coefficient of the Pd(II)-3,4-DHBINH complex were 1.08 and 0.04 respectively. Major cations and anions did not show any interference. Anti-microbial activity of the Pd(II)-3,4-DHBINH has been studied. The developed method has been successfully applied to the analysis of Pd(II) in spiked samples. Comparing the results with those obtained using an atomic absorption spectrophotometer tested the validity of the method

Interaction of D-phenylalanine with Co(II)-substituted rabbit muscle pyruvate kinase: kinetic and optical properties

1982 ◽  
Vol 60 (9) ◽  
pp. 861-866
Author(s):  
Chiu-Yin Kwan ◽  
Robert C. Davis
Keyword(s):  
Amino Acids ◽  
Optical Properties ◽  
Pyruvate Kinase ◽  
Conformational Changes ◽  
Metal Ion ◽  
Inhibitory Effect ◽  
Rabbit Muscle ◽  
Divalent Metal Ion ◽  
Muscle Pyruvate Kinase ◽  
Difference Absorption

The kinetic and optical properties of Co(II)-substituted pyruvate kinase in the presence of D-phenylalanine (D-Phe) were investigated. The results are discussed in comparison with the effects of its optical isomer L-phenylalanine (L-Phe) on the same enzyme. The catalytic effect of D-Phe on rabbit muscle pyruvate kinase depended upon the nature of the activating divalent metal ion used. It has stimulatory effect on Mg(II)-activated enzyme, but inhibitory effect on Co(II)-activated enzyme. Unlike the inhibitory effect of L-Phe, the inhibition of Co(II)–enzyme by D-Phe was not sensitive to the changes of pH and temperature, could not be reversed by L-alanine (L-Ala), displayed hyperbolic kinetics, and was noncompetitive with respect to phosphoenolpyruvate saturation. D-Phe induced substantial visible circular dichroism (CD) spectral changes of Co(II)–enzyme similar to those induced by L-Phe. Although ultraviolet CD spectrum was not affected, D-Phe induced an ultraviolet difference absorption spectral change very similar to, but much smaller than, that induced by L-Phe. Our results support that D-Phe and other amino acids interact with the enzyme at two different sites: a common site, causing similar conformational changes which bear little direct kinetic relevance, and a kinetically relevant site, which is sterically dependent upon the side chain of the amino acids.

Structural basis for the allosteric regulation of the SbtA bicarbonate transporter by the PII-like protein, SbtB, from Cyanobium sp. PCC7001

2019 ◽  
Author(s):  
Joe A. Kaczmarski ◽  
Nan-Sook Hong ◽  
Bratati Mukherjee ◽  
Laura T. Wey ◽  
Loraine Rourke ◽  
...  
Keyword(s):  
Metal Ion ◽  
Transcriptional Control ◽  
Allosteric Regulation ◽  
Inhibitory Effect ◽  
Photosynthetic Performance ◽  
Regulatory Proteins ◽  
Structural Basis ◽  
Titration Calorimetry ◽  
X Ray ◽  
Bicarbonate Transporter

ABSTRACTCyanobacteria have evolved a suite of enzymes and inorganic carbon (Ci) transporters that improve photosynthetic performance by increasing the localized concentration of CO2 around the primary CO2-fixating enzyme, Rubisco. This CO2-concentrating mechanism (CCM) is highly regulated, responds to illumination/darkness cycles and allows cyanobacteria to thrive under limiting Ci conditions. While the transcriptional control of CCM activity is well understood, less is known about how regulatory proteins might allosterically regulate Ci transporters in response to changing conditions. Cyanobacterial sodium-dependent bicarbonate transporters (SbtAs) are inhibited by PII-like regulatory proteins (SbtBs), with the inhibitory effect being modulated by adenylnucleotides. Here, we used isothermal titration calorimetry to show that SbtB from Cyanobium sp. PCC7001 (SbtB7001) binds AMP, ADP, cAMP and ATP with micromolar-range affinities. X-ray crystal structures of apo- and nucleotide-bound SbtB7001 revealed that while AMP, ADP and cAMP have little effect on the SbtB7001 structure, binding of ATP stabilizes the otherwise flexible T-loop and that the flexible C-terminal C-loop adopts several distinct conformations. We also show that ATP binding affinity is increased ten-fold in the presence of Ca2+ and we present an X-ray crystal structure of Ca2+ATP:SbtB7001 that shows how this metal ion facilitates additional stabilizing interactions with the apex of the T-loop. We propose that the Ca2+ATP-induced conformational change observed in SbtB7001 is important for allosteric regulation of SbtA activity by SbtB and is consistent with changing adenylnucleotide levels in illumination/darkness cycles.GRAPHICAL ABSTRACT

Complete genome sequence of Micromonospora craniellae LHW63014T, a potential metal ion-chelating agent producer

2020 ◽  
pp. 100830
Author(s):  
Xue Yang ◽  
Zhong-Hui Zhu ◽  
Xia Ji ◽  
Zhao-Ming Liu ◽  
Hua Zhang ◽  
...  
Keyword(s):  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Metal Ion ◽  
Chelating Agent

scholarly journals Studies on Analogues of L-Cysteine and L-Cystine

Blood ◽  
1956 ◽  
Vol 11 (1) ◽  
pp. 1-10 ◽  
Author(s):  
AUSTIN S. WEISBERGER ◽  
LEIF G. SUHRLAND ◽  
JOSEPH SEIFTER
Keyword(s):  
Amino Acids ◽  
Spatial Configuration ◽  
Spatial Relationship ◽  
Inhibitory Effect ◽  
Selenium Compounds ◽  
Low Concentrations ◽  
Relationship Of ◽  
Normal Cellular Function

Abstract The amino acids L-cysteine and L-cystine appear to have an important role in the metabolism of leukocytes. Decreased availability of these amino acids may therefore have important effects on leukocytes. The possibility of decreasing the influx of radioactive L-cystine into leukemic leukocytes was investigated by exposing the leukocytes to various analogues of cysteine (cystine) prior to incubation with S35 L-cystine. It was found that a highly specific structural and spatial configuration is required to decrease the influx of S35 L-cystine. Thus unlabeled L-cysteine is effective in decreasing the incorporation of radioactive L-cystine. However, analogues of cystine in which there is modification or substitution of the sulfhydryl, amino or carboxyl group do not decrease the influx of S35 L-cystine. Furthermore, any alteration in the spatial relationship of the sulfhydryl and amino groups of L-cysteine also results in a loss of the ability of an analogue to decrease the incorporation of S35 L-cystine. Of the compounds studied and in the concentrations employed, only unlabeled L-cysteine, selenium cystine and phenyl selenium cysteine were effective. Selenium cystine is identical with cystine except that selenium replaces the sulfur in the molecule. Phenyl selenium cysteine is also closely related structurally to cysteine. The mechanism of action of selenium cystine and phenyl selenium cysteine in decreasing the influx of S35 L-cystine is not known. Other selenium compounds tested were ineffective. These compounds may exert their inhibitory effect by (a) competitive combination with specific intracellular receptors for L-cysteine (L-cystine), (b) inactivation of enzymes or compounds essential for normal cellular function, (c) alteration in membrane permeability or (d) a toxic effect of selenium. Since selenium cystine and phenyl selenium cystine are inhibitory in low concentrations in vitro, these compounds may have important effects on leukemic leukocytes in vivo.

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