Activation of the phosphosignaling protein CheY. I. Analysis of the phosphorylated conformation by 19F NMR and protein engineering

CheY, the 14-kDa response regulator protein of the Escherichia coli chemotaxis pathway, is activated by phosphorylation of Asp57. In order to probe the structural changes associated with activation, an approach which combines 19F NMR, protein engineering, and the known crystal structure of one conformer has been utilized. This first of two papers examines the effects of Mg(II) binding and phosphorylation on the conformation of CheY. The molecule was selectively labeled at its six phenylalanine positions by incorporation of 4-fluorophenylalanine, which yielded no significant effect on activity. One of these 19F probe positions monitored the vicinity of Lys109, which forms a salt bridge to Asp57 in the apoprotein and has been proposed to act as a structural "switch" in activation. 19F NMR chemical shift studies of the labeled protein revealed that the binding of the cofactor Mg(II) triggered local structural changes in the activation site, but did not perturb the probe of the Lys109 region. The structural changes associated with phosphorylation were then examined, utilizing acetyl phosphate to chemically generate phsopho-CheY during NMR acquisition. Phosphorylation triggered a long-range conformational change extending from the activation site to a cluster of 4 phenylalanine residues at the other end of the molecule. However, phosphorylation did not perturb the probe of Lys109. The observed phosphorylated conformer is proposed to be the first step in the activation of CheY; later steps appear to perturb Lys109, as evidenced in the following paper. Together these results may give insight into the activation of other prokaryotic response regulators

"Hot and sticky" and "cold and damp" pyroclastic eruptions, and their relationship with topography: valley- and lake-filling ignimbrites, Ardnamurchan, NW Scotland

Peer reviewe

Exact relativistic stellar models with liquid surface. I. Generalizing Buchdahl's n=1n=1 polytrope

A family of exact relativistic stellar models is described. The family generalizes Buchdahl's n=1 polytropic solution. The matter content is a perfect fluid and, excluding Buchdahl's original model, it behaves as a liquid at low pressures in the sense that the energy density is non-zero in the zero pressure limit. The equation of state has two free parameters, a scaling and a stiffness parameter. Depending on the value of the stiffness parameter the fluid behaviour can be divided in four different types. Physical quantities such as masses, radii and surface redshifts as well as density and pressure profiles are calculated and displayed graphically. Leaving the details to a later publication, it is noted that one of the equation of state types can quite accurately approximate the equation of state of real cold matter in the outer regions of neutron stars. Finally, it is observed that the given equation of state does not admit models with a conical singularity at the center.Comment: 19 pages, 12 figures (16 eps files), LaTeX2e with the standard packages amssymb, amsmath, graphicx, subfigure, psfra

ROSAT HRI observations of the intermediate-age open cluster IC 4756

We have obtained an 88 kilosecond ROSAT HRI exposure of the intermediate-age open cluster IC 4756 with the purpose of detecting stars in the high luminosity tail (log LX ≥1029 erg s−1) of its X-ray luminosity distribution. However, only 1 cluster member (HSS 201) out of the 60 members inside the central high-sensitivity region of the HRI field of view (FOV) was detected. This star has spectral type A8, suggesting a close binary system with a low mass X-ray emitting companion. We compare the distribution of upper limits for F and G-type dwarfs in IC 4756 with the X-ray distribution functions of the similarly aged Hyades and Praesepe clusters. The results of this statistical analysis are inconclusive for G-type stars, but suggest that at least F-type stars in IC 4756 are not as X-ray luminous as their Hyades counterparts, thus indicating intrinsic differences between the two clusters. Finally, our data indicate a deficit of very active binaries with respect to both Hyades and Praesepe, and older open clusters

Discrete choice modelling for traffic densities with lane-change behaviour

This paper investigates the modelling for traffic densities with lane-change behaviour using the information provided by loop detectors. The existing studies on traffic density estimation for multi-lane roadways mainly focus on the scenario where either vehicles’ lane-change manoeuvres are not common or the lane-change pattern is time-invariant. This research, however, takes into consideration the time-varying nature of drivers’ lane-change manoeuvres, and models the lane-change probabilities using a number of discrete choice models. These lane-change models are then embedded into a state space model to capture the dynamics of traffic flow. The extended Kalman filter is used to update the estimated traffic densities of multi-lane motorways. A numerical study is carried out to investigate the performance of the developed approach