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Site-specific and synergistic stimulation of methylation on the bacterial chemotaxis receptor Tsr by serine and CheW
BACKGROUND: Specific glutamates in the methyl-accepting chemotaxis proteins (MCPs) of Escherichia coli are modified during sensory adaptation. Attractants that bind to MCPs are known to increase the rate of receptor modification, as with serine and the serine receptor (Tsr), which contributes to an increase in the steady-state (adapted) methylation level. However, MCPs form ternary complexes with two cytoplasmic signaling proteins, the kinase (CheA) and an adaptor protein (CheW), but their influences on receptor methylation are unknown. Here, the influence of CheW on the rate of Tsr methylation has been studied to identify contributions to the process of adaptation. RESULTS: Methyl group incorporation was measured in a series of membrane samples in which the Tsr molecules were engineered to have one available methyl-accepting glutamate residue (297, 304, 311 or 493). The relative rates at these sites (0.14, 0.05, 0.05 and 1, respectively) differed from those found previously for the aspartate receptor (Tar), which was in part due to sequence differences between Tar and Tsr near site four. The addition of CheW generated unexpectedly large and site-specific rate increases, equal to or larger than the increases produced by serine. The increases produced by serine and CheW (added separately) were the largest at site one, ~3 and 6-fold, respectively, and the least at site four, no change and ~2-fold, respectively. The rate increases were even larger when serine and CheW were added together, larger than the sums of the increases produced by serine and CheW added separately (except site four). This resulted in substantially larger serine-stimulated increases when CheW was present. Also, CheW enhanced methylation rates when either two or all four sites were available. CONCLUSION: The increase in the rate of receptor methylation upon CheW binding contributes significantly to the ligand specificity and kinetics of sensory adaptation. The synergistic effect of serine and CheW binding to Tsr is attributed to distinct influences on receptor structure; changes in the conformation of the Tsr dimer induced by serine binding improve methylation efficiency, and CheW binding changes the arrangement among Tsr dimers, which increases access to methylation sites
Solid-State NMR Demonstrates that Active Signaling Complexes of Bacterial Chemoreceptors Do Not Adopt the Proposed Trimer-Of-Dimers Structure
Scaled Chrysophyceae From Lake Itasca Region. ll. Synura, Chrysophaerella, Spiniferomonas
Using electron microscopy, 49 plankton samples from the Lake Itasca region were examined for the silica-scaled chrysophycean genera Synura, Chrysosphaerella and Spiniferomonas. Twelve taxa were observed: five are new for Minnesota, and two of these, Synura multidenta and Synura petersenil f. asmundiae, are new reports for the continental United States
Scaled Chrysophycae From Lake Itasca Region I. Mallomonas
By means of electron microscopy, phytoplankton samples from the Lake Itasca region were examined for the silica-scaled chrysophycean genus Mallomonas. Seventeen taxa were observed: 15 are new for Minnesota, of these 15, seven are also new reports for the continental United States
Optimization and performance of an optical cardio-magnetometer
Cardiomagnetometry is a growing field of noninvasive medical diagnostics that
has triggered a need for affordable high-sensitivity magnetometers. Optical
pumping magnetometers are promising candidates satisfying that need since it
was demonstrated that they can map the heart magnetic field. For the
optimization of such devices theoretical limits on the performance as well as
an experimental approach is presented. The promising result is a intrinsic
magnetometric sensitivity of 63 fT / Hz^1/2 a measurement bandwidth of 140 Hz
and a spatial resolution of 28 mm
Pachycormid fish fed on octobrachian cephalopods: new evidence from the ‘Schistes bitumineux’ (early Toarcian) of southern Luxembourg
A re-examination of the early Toarcian fish fossils preserved in public paleontological collections in Luxembourg revealed 70 specimens of large Toarcian pachycormid fish with an excellent three-dimensional preservation within calcareous nodules. Six of them are associated with octobrachian coleoid gladii in their oesophagus or stomach, an association not previously described from Luxembourg. The pachycormids are ascribed to Pachycormus macropterus (Blainville, 1818) and Saurostomus esocinus Agassiz, 1843 while the octobrachian gladii are ascribed to Teudopsis bollensis Voltz, 1836, Teudopsis sp. indet. and Loligosepiidae indet. The position and orientation of the gladii provide direct evidence of these fishes feeding on coleoids and thus a teuthophagous diet, rather than an accidental joint burial. Together with evidence from coeval deposits in Germany, these findings suggest that teuthophagy was a widespread feeding strategy at the base of the clade that contains the suspension-feeding pachycormid giants of the Jurassic–Cretaceous
Ideology In The Evolution of Nonprofit Academic Programs
How do academic programs evolve? This overarching question has several subcomponents such as the following: 1. What gives impetus to new programs on a campus? 2. Do new programs begin as undergraduate majors and degrees that eventually progress to the graduate level? 3. Are there critical faculty masses that are necessary for the evolution of programs? These and similar questions are addressed in this analysis of four case studies examined over a 20 plus year period. Guiding this inquiry is the literature on organizational development as well as academic program development and sustainability. Among the findings are these: 1. External and internal factors of a campus including beliefs or ideologies influence the development of new programs. 2. While one might think that undergraduate level programs precede graduate programs on a campus, programs do not necessarily develop in this manner. 3. Critical faculty levels may be more myth than reality. These conclusions and others from this inquiry may be common sense on one level but the reality is that they are not. The purpose of this inquiry is to debunk often-embraced myths in higher education
Optimization and Structural Stability of Gold Nanoparticle–Antibody Bioconjugates
Gold nanoparticles (AuNPs) bound with biomolecules have emerged as suitable biosensors exploiting unique surface chemistries and optical properties. Many efforts have focused on antibody bioconjugation to AuNPs resulting in a sensitive bioconjugate to detect specific types of bacteria. Unfortunately, bacteria thrive under various harsh environments, and an understanding of bioconjugate stability is needed. Here, we show a method for optimizing Listeria monocytogenes polyclonal antibodies bioconjugation mechanisms to AuNPs via covalent binding at different pH values, from 2 to 11, and 2-(N-morpholino)ethanesulfonic acid (MES), 3-(N-morpholino)propanesulfonic acid, NaOH, HCl conditions. By fitting Lorentz curves to the amide I and II regions, we analyze the stability of the antibody secondary structure. This shows an increase in the apparent breakdown of the antibody secondary structure during bioconjugation as pH decreases from 7.9 to 2. We find variable adsorption efficiency, measured as the percentage of antibody adsorbed to the AuNP surface, from 17 to 27% as pH increases from 2 to 6 before decreasing to 8 and 13% at pH 7.9 and 11, respectively. Transmission electron microscopy (TEM) analysis reveals discrepancies between size and morphological changes due to the corona layer assembly from antibody binding to single nanoparticles versus aggregation or cluster self-assembly into large aggregates. The corona layer formation size increases from 3.9 to 5.1 nm from pH 2 to 6, at pH 7.9, there is incomplete corona formation, whereas at pH 11, there is a corona layer formed of 6.4 nm. These results indicate that the covalent binding process was more efficient at lower pH values; however, aggregation and deactivation of the antibodies were observed. We demonstrate that optimum bioconjugation condition was determined at pH 6 and MES buffer-type by indicators of covalent bonding and stability of the antibody secondary structure using Fourier transform-infrared, the morphological characteristics and corona layer formation using TEM, and low wavelength shifts of ultraviolet–visible after bioconjugation
Mechanisms and drivers of belemnite body-size dynamics across the Pliensbachian–Toarcian crisis
Body-size reduction is considered an important response to current climate warming and has been observed during past biotic crises, including the Pliensbachian–Toarcian crisis, a second-order mass extinction. However, in fossil cephalopod studies, the mechanisms and their potential link with climate are rarely investigated and palaeobiological scales of organization are not usually differentiated. Here, we hypothesize that belemnites reduce their adult size across the Pliensbachian–Toarcian boundary warming event. Belemnite body-size dynamics across the Pliensbachian–Toarcian boundary in the Peniche section (Lusitanian Basin, Portugal) were analysed based on the newly collected field data. We disentangle the mechanisms and the environmental drivers of the size fluctuations observed from the individual to the assemblage scale. Despite the lack of a major taxonomic turnover, a 40% decrease in rostrum volume is observed across the Pliensbachian–Toarcian boundary, before the Toarcian Oceanic Anoxic Event where belemnites go locally extinct. The pattern is mainly driven by a reduction in adult size of the two dominant species, Pseudohastites longiformis and Passaloteuthis bisulcata. Belemnite-size distribution is best correlated with fluctuations in a palaeotemperature proxy (stable oxygen isotopes); however, potential indirect effects of volcanism and carbon cycle perturbations may also play a role. This highlights the complex interplay between environmental stressors (warming, deoxygenation, nutrient input) and biotic variables (productivity, competition, migration) associated with these hyperthermal events in driving belemnite body-size
Refining the marine reptile turnover at the Early–Middle Jurassic transition
peer reviewedEven though a handful of long-lived reptilian clades dominated Mesozoic marine ecosystems, several biotic turnovers drastically changed the taxonomic composition of these communities. A seemingly slow paced, within-geological period turnover took place across the Early-Middle Jurassic transition. This turnover saw the demise of early neoichthyosaurians, rhomaleosaurid plesiosaurians and early plesiosauroids in favour of ophthalmosaurid ichthyosaurians and cryptoclidid and pliosaurid plesiosaurians, clades that will dominate the Late Jurassic and, for two of them, the entire Early Cretaceous as well. The fossil record of this turnover is however extremely poor, and this change of dominance appears to be spread across the entire middle Toarcian-Bathonian interval.
We describe a series of ichthyosaurian and plesiosaurian specimens from successive geological formations in Luxembourg and Belgium that detail the evolution of marine reptile assemblages across the Early-Middle Jurassic transition within a single area, the Belgo-Luxembourgian sub-basin. These fossils reveal the continuing dominance of large rhomaleosaurid plesiosaurians, microcleidid plesiosaurians, and Temnodontosaurus-like ichthyosaurians up to the latest Toarcian, indicating that the structuration of the upper tier of Western Europe marine ecosystems remained essentially constant up to the very end of the Early Jurassic. These fossils also suddenly record ophthalmosaurid ichthyosaurians and cryptoclidid plesiosaurians by the early Bajocian. These results from a geographically-restricted area provide a clearer picture of the shape of the marine reptile turnover occurring at the early–Middle Jurassic transition. This event appears restricted to the sole Aalenian stage, reducing the uncertainty of its duration, at least for ichthyosaurians and plesiosaurians, to 4 instead of 14 million years
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