From the gating charge response to pore domain movement: Initial motions of Kv1.2 dynamics under physiological voltage changes

Abstract

Recent structures of the potassium channel provide an essential beginning point for explaining how the pore is gated between open and closed conformations by changes in membrane voltage. Yet, the molecular details of this process and the connections to transmembrane gradients are not understood. To begin addressing how changes within a membrane environment lead to the channel’s ability to sense shifts in membrane voltage and to gate, we performed double-bilayer simulations of the Kv1.2 channel. These double-bilayer simulations enable us to simulate realistic voltage drops from resting potential conditions to depolarized conditions by changes in the bath conditions on each side of the bilayer. Our results show how the voltage sensor domain movement responds to differences in transmembrane potential. The initial voltage sensor domain movement, S4 in particular, is modulated by the gating charge response to changes in voltage and is initially stabilized by the lipid headgroups. We show this response is directly coupled to the initial stages of pore domain motion. Results presented here provide a molecular model for how the pre-gating process occurs in sequential steps: Gating charge response, movement and stabilization of the S4 voltage sensor domain, and movement near the base of the S5 region to close the pore domain.     

Beneficial effects of n-hexane bark extract of Onosma echioides L. on diabetic peripheral neuropathy

Onosma echioides Linn (Boraginaceae) is the most frequently used curative herb widely used for kidney obstruction, sciatic pain, and gout. The present study was designed to investigate the therapeutic effects of n-hexane bark extract of O. echioides (OE) L. root in vivo against Streptozotocin-induced diabetic neuropathy in SD rats. For in vivo activity, the experiment was categorized into five different groups (n = 5). Group-I was considered as nondiabetic/normal control (NC) treated with 0.5% carboxymethyl cellulose (CMC), Group II as diabetic control, Group-III, IV, and V served as diabetic treated with OE 50, OE 100, and pregabalin at a dose of 50, 100, and 10 mg/kg body weight, orally, respectively. Body weight, blood glucose, oral glucose tolerance test, behavioral studies (motor coordination test, thermal hyperalgesia, cold allodynia, locomotor activity, oxidative biomarkers (thio barbituric acid reactive substances [TBARS], superoxide dismutase [SOD], glutathione [GSH], and catalase), and histopathology of the sciatic nerve were performed. Treatment with OE showed a dose-dependent increase in neuroprotective activity by improving the myelination and decreasing the axonal swelling of nerve fibers. The verdicts of behavioral activities showed a remarkable effect on animals after the treatment of extract and standard drug pregabalin. In conclusion, our findings supported the traditional application of OE and explored its importance in the management of diabetic neuropathy. Additional clinical experiments may provide novel therapeutic drugs for diabetes and its complications.     

Synthesis of 5‐tert‐butyl‐1‐(3‐tert‐butyldimethylsiloxy)phenyl‐4,4‐dimethyl‐2,6,7‐trioxabicyclo[3.2.0]heptanes and their fluoride‐induced chemiluminescent decomposition: effect of a phenolic electron donor on the CIEEL decay rate in aprotic polar solvent

Four bicyclic dioxetanes bearing a phenolic substituent, 3‐tert‐butyldimethylsiloxy‐4‐chlorophenyl ( 3a ), 5‐tert‐butyldimethylsiloxy‐4‐chloro‐2‐ethylphenyl ( 3b ), 5‐tert‐butyldimethylsiloxy‐2‐ethylphenyl ( 3c ), and 3‐tert‐butyldimethylsiloxy‐4‐ethylphenyl ( 3d ), were synthesized. All dioxetanes 3a – 3d gave intense blue light on treatment with tetrabutylammonium fluoride (TBAF) in DMSO or acetonitrile. Kinetic study on the fluoride‐induced CIEEL decay of these dioxetanes 3a – 3d and the parent dioxetane 2b revealed that the para‐substitution with chlorine on the phenolic moiety of dioxetane increases free energy of activation (ΔG?), while the para‐substitution with ethyl on the aryl decreases ΔG?. On the other hand, substitution with an ethyl at the ortho‐position instead of the para‐position was found to increase ΔG? and to suppress the CIEEL decay. This fact is attributed to the steric factor of the ortho‐ethyl group which would prevent the aromatic ring from rotating freely around the axis joined to the peroxide ring, and supports the suggestion for a CIEEL‐active dioxetane bearing a phenolic moiety that an intramolecular electron transfer occurs preferentially from the phenolic donor to O–O of the dioxetane ring, when the aromatic ring lies in a certain conformation(s). Copyright © 2002 John Wiley & Sons, Ltd.     

Scale-dependent interactions between intrinsic and extrinsic processes reduce variability in protist populations

Interactions between intrinsic processes and extrinsic fluctuations can positively impact population persistence in ways often not predicted by classic ecological models. These interactions only arise when the intrinsic and extrinsic processes operate on the proper relative scales in time or space. Both metapopulation theory and resonance/attenuation theory suggest that interactions which lower population variability will occur when the intrinsic and extrinsic process occur on similar time scales. I performed an aquatic protist microcosm experiment to investigate how the relative frequencies of extrinsic density perturbations and intrinsic resource pulses impacted population variability. Population variability was lowest in the treatments of intermediate frequency, in which the extrinsic fluctuations and intrinsic processes were on the same time scale. This result is consistent with general theoretical predictions, and empirically documents the importance of considering scale in interactions between intrinsic and extrinsic processes that positively impact population persistence.     

Effects of an inhomogeneous magnetic field on flowing erythrocytes

Effects of an inhomogeneous magnetic field on narrow erythrocyte streams in a wide and transparent laminar buffer flow were studied. The stream line of erythrocytes containing paramagnetic hemoglobin showed distinct displacement toward the stronger magnetic field. The displacement increased in the order, oxygenated erythrocytes (no displacement), erythrocytes containing cyanomethemoglobin, deoxygenated erythrocytes, erythrocytes containing methemoglobin in the high spin state; more precisely the displacement was proportional to the square of the paramagnetic moment of hemoglobin contained in the erythrocytes. In addition, the displacement was proportional to the product of the magnetic flux density and its gradient, and approximately proportional to the hematocrit of the flowing-erythrocyte suspension, and was much larger than that calculated for a single erythrocyte. These phenomena could be successfully interpreted by the interaction of paramagnetic erythrocytes with the inhomogeneous magnetic field, the resistance force (Stokes Law) from the bulk water, and the hydrodynamic interaction between erythrocytes.     

Chick brain glutamine synthetase and Mn2+−Mg2+ interactions

Glutamine synthetase (GS) from the chick brain was purified to apparent homogeneity by ammonium sulfate fractionation followed by affinity chromatography, electrofocusing and Sephadex G-150 chromatography. The purified enzyme showed a single band on sodium dodecyl sulfate analysis in polyacrylamide gel. By sedimentation equilibrium analysis and gel electrophoresis analysis, it was shown that the enzyme has a subunit molecular weight of 45,000 and a native molecular weight of 364,000, which is consistent with an octameric structure. Sedimentation analysis in the presence of Mg²⁺ revealed three different forms of macromolecules corresponding respectively to a monomer, a tetramer and an octamer. Among eight cations tested (Ca²⁺, Co²⁺, Fe²⁺, Li⁺, Mg²⁺, Mn²⁺, Ni²⁺, Zn²⁺) only Co²⁺, Mg²⁺ and Mn²⁺ supported GS activity; the order of activatory ability was Mg²⁺>Co²⁺>Mn²⁺. The maximum activating effect of Mn²⁺ occurs only within a very narrow range of concentration: with an excess of cation causing strong inhibition of GS activity. For each cation, maximal GS activity occurs at a defined cation/ATP ratio. A regulatory system in which Mn²⁺, modulates the Mg²⁺ dependent GS activity, is proposed; such cation interactions may be of significance in the intracellular control of glutamine synthesis.     

A locus involved in kanamycin, chloramphenicol and L-serine resistance is located in the bglY-galU region of the Escherichia coli K12 chromosome

Summary Spontaneous mutants of Escherichia coli K12 displaying an increased level of the kanamycin resistance conferred by plasmid pGR71 were selected. Several mutants obtained in this way apparently carry large chromosomal deletions extending into galU and/or bglY (27 min). This positive selection of deletions allowed detection of a new locus located between galU and bglY. Deletions of this locus are responsible for increased resistance to kanamycin (Irk), decreased resistance to l-serine in minimal medium (Drs) and decreased resistance to chloramphenicol (Drc) when a cat gene is present in the bacteria.     

Morphometric comparison of populations of Orchis sitnia Lam. (Orchidaceae) from Oxfordshire and Kent

BATEMAN, R. M. & FARRINGTON, O. S., 1989. Morphometric comparison of populations of Orchis sitnia Lam. (Orchidaceae) from Oxfordshire and Kent. Univariate and multivariate morphometric analyses of Orchis simia Lam. from its two remaining naturally-founded populations in Britain (Goring, Oxfordshire and Faversham, Kent) reveal only minor morphological differences between the populations and suprisingly low levels of intra-population variation. This probably reflects recent founding from Continental seed at Faversham and penecontemporaneous, human-induced, catastrophic contraction of the gene pool at Goring. The apparently restricted gene pools of these populations may adversely effect their long-term survival.     

The “ricochet effect” and prey capture in colonial spiders

Summary Increased prey capture efficiency in colonial spiders is a consequence of the ricochet effect, as prey are captured after they bounce off several webs in succession. In this study, the prey capture of three species of colonial spiders in the genus Metepeira from Mexico are compared. These species, from different habitats, show varying levels of social organization (group size and withingroup spacing) that affect prey capture from ricochets. Metepeira sp. a (a presumed new species tentatively named atascadero) from desert grassland habitats, occur solitarily or in small groups, and gain little from prey ricochets: prey capture rates are low and variance in prey captured/spider is high. M. spinipes, from mesic agricultural sites, occur in groups of 10–150, and show a ricochet effect resulting in more and larger prey, and reduced variance in capture rate. M. incrassata, from tropical rainforest/agricultural sites, occur in large colonies of hundreds to thousands of individuals, and show a similar ricochet effect. The ricochet effect does not influence taxonomic composition of prey in either M. atascadero or M. spinipes, but does in tropical M. incrassata. This result, however, is primarily due to the capacity of certain taxa (eg., Lepidoptera), more common in the tropics, to escape more easily from spider webs. A comparison of prey capture efficiency of colonial M. incrassata with that of solitary M. atascadero shows that the ricochet effect provides an increase in efficiency across all size classes of prey.