Proteomic analysis of resting and thrombin-stimulated platelets reveals the translocation and functional relevance of HIP-55 in platelets

The platelet surface is a dynamic interface that changes rapidly in response to stimuli to coordinate the formation of thrombi at sites of vascular injury. Tight control is essential as loss of organisation may result in the inappropriate formation of thrombi (thrombosis) or excessive bleeding. In this paper we describe the comparative analysis of resting and thrombin-stimulated platelet membrane proteomes and associated proteins to identify proteins important to platelet function. Surface proteins were labelled using a biotin tag and isolated by NeurtrAvidin affinity chromatography. Liquid phase IEF and SDS-PAGE were used to separate proteins, and bands of increased intensity in the stimulated platelet fractions were digested and identified by FT-ICR mass spectrometry. Novel proteins were identified along with proteins known to be translocated to the platelet surface. Furthermore, many platelet proteins revealed changes in location associated with function, including G6B and Hip-55. HIP-55 is an SH3-binding protein important in T-cell receptor signalling. Further analysis of HIP-55 revealed that this adaptor protein becomes increasingly associated with both Syk and integrin beta 3 upon platelet activation. Analysis of HIP-55 deficient platelets revealed reduced fibrinogen binding upon thrombin stimulation, suggesting HIP-55 to be an important regulator of platelet function.

The structure of echovirus type 12 bound to a two-domain fragment of its cellular attachment protein decay-accelerating factor (CD 55)

Echovirus type 12 (EV12), an enterovirus of the Picornaviridae family, uses the complement regulator, decay-accelerating factor (DAF, CD55) as a cellular receptor. We have calculated a three-dimensional reconstruction of EV12 bound to a fragment of DAF, consisting of short consensus repeat domains 3 and 4, from cryo-negative stain electron microscopy data (EMD #1057). This shows that, as for an earlier reconstruction of the related echovirus type 7 bound to DAF, attachment is not within the viral canyon but occurs close to the two-fold symmetry axes. Despite this general similarity, our reconstruction reveals a receptor interaction that is quite different from that observed for EV7. Fitting of the crystallographic co-ordinates for DAF34 and EV11 into the reconstruction shows a close agreement between the crystal structure of the receptor fragment and the density for the virus-bound receptor, allowing unambiguous positioning of the receptor with respect to the virion (PDB #1UPN). Our finding that the mode of virus-receptor interaction in EV12 is distinct from that seen for EV7 raises interesting questions regarding the evolution and biological significance of the DAF-binding phenotype in these viruses.

Flow in the high latitude ionosphere: measurements at 15s resolution made using the EISCAT ‘Polar’ experiment

We present a first overview of flows in the high latitude ionosphere observed at 15 s resolution using the U.K.-Polar EISCAT experiment. Data are described from experiments conducted on two days, 27 October 1984 and 29 August 1985, which together span the local times between about 0200 and 2130MLT and cover five different regions of ionospheric flow. With increasing local time, these are: the dawn auroral zone flow cell, the dayside region of low background flows equatorward of the flow cells, the dusk auroral zone flow cell, the boundary region between the dusk auroral zone and the polar cap, and the evening polar cap. Flows in both the equatorward and poleward portions of the auroral zone cells appear to be relatively smooth, while in the central region of high speed flow considerable variations are generally present. These have the form of irregular fluctuations on a wide range of time scales in the early morning dawn cell, and impulsive wave-like variations with periods of a few minutes in the afternoon dusk cell. In the dayside region between the flow cells, the ionosphere is often essentially stagnant for long intervals, but low amplitude ULF waves with a period of about 5 min can also occur and persist for many cycles. These conditions are punctuated at one to two hour intervals by sudden ‘flow burst’ events with impulsively generated damped wave trains. Initial burst flows are generally directed poleward and can peak at line-of-sight speeds in excess of 1 km s^{−1} after perhaps 45 s. Flows in the polar cap are reasonably smooth on time scales of a few minutes and show no evidence for the presence of ULF waves. Under most, but not all, of the above conditions, the beam-swinging algorithm used to determine background vector flows should produce meaningful results. Comparison of these flow data with simultaneous plasma and magnetic field measurements in the solar wind, made by the AMPTE IRM and UKS spacecraft, emphasizes the strong control exerted on high latitude flows by the north-south component of the IMF.

Climatology of polar lows over the Sea of Japan using the JRA-55 reanalysis

Polar lows are intense meso- a -scale cyclones that develop over the oceans poleward of the main baroclinic zone. A number of previous studies have reported polar low formation over the Sea of Japan within the East Asian winter monsoon. To understand the climatology of polar lows over the Sea of Japan, a tracking al- gorithm for polar lows is applied to the recent JRA-55 reanalysis. The polar low tracking is applied to 36 cold seasons (October–March) from October 1979 to March 2015. The polar lows over the Sea of Japan reach their maximum intensity on the southeastern side of the midline between the Japanese islands and the Asian continent. Consistent with previous case studies, composite analysis demonstrates that the polar low devel- opment is associated with the enhanced northerly flow on the western side of a synoptic-scale extratropical cyclone, with the cold trough in the midtroposphere and with increased heat fluxes from the sea surface. Furthermore, the present climatological study has revealed two dominant directions of motion of the polar lows: southward and eastward. Southward-moving polar lows are steered by a strong northerly flow in the lower troposphere, which is enhanced on the western side of synoptic-scale extratropical cyclones, while the eastward-moving polar lows occur within a planetary-scale westerly flow in the midlatitudes. Thus, the di- rection of polar low motion reflects the difference in planetary- and synoptic-scale conditions.