Src homology 2 domain containing protein 5 (SH2D5) binds the breakpoint cluster region protein, BCR, and regulates levels of Rac1-GTP

SH2D5 is a mammalian-specific, uncharacterized adaptor-like protein that contains an N-terminal phosphotyrosine binding (PTB) domain and a C-terminal Src Homology 2 (SH2) domain. We show that SH2D5 is highly enriched in adult mouse brain, particularly in purkinjie cells in the cerebellum and the cornu ammonis of the hippocampus. Despite harboring two potential phosphotyrosine (pTyr) recognition domains, SH2D5 binds minimally to pTyr ligands, consistent with the absence of a conserved pTyr-binding arginine residue in the SH2 domain. Immunoprecipitation coupled to mass spectrometry (IP-MS) from cultured cells revealed a prominent association of SH2D5 with Breakpoint Cluster Region protein (BCR), a RacGAP that is also highly expressed in brain. This interaction occurred between the PTB domain of SH2D5 and an NxxF motif located within the N-terminal region of BCR. siRNA-mediated depletion of SH2D5 in a neuroblastoma cell line, B35, induced a cell rounding phenotype correlated with low levels of activated Rac1-GTP, suggesting that SH2D5 affects Rac1-GTP levels. Taken together, our data provide the first characterization of the SH2D5 signaling protein

Vortex Dynamics and Defects in Simulated Flux Flow

We present the results of molecular dynamic simulations of a two-dimensional vortex array driven by a uniform current through random pinning centers at zero temperature. We identify two types of flow of the driven array near the depinning threshold. For weak disorder the flux array contains few dislocation and moves via correlated displacements of patches of vortices in a {\it crinkle} motion. As the disorder strength increases, we observe a crossover to a spatially inhomogeneous regime of {\it plastic} flow, with a very defective vortex array and a channel-like structure of the flowing regions. The two regimes are characterized by qualitatively different spatial distribution of vortex velocities. In the crinkle regime the distribution of vortex velocities near threshold has a single maximum that shifts to larger velocities as the driving force is increased. In the plastic regime the distribution of vortex velocities near threshold has a clear bimodal structure that persists upon time-averaging the individual velocities. The bimodal structure of the velocity distribution reflects the coexistence of pinned and flowing regions and is proposed as a quantitative signature of plastic flow.Comment: 12 pages, 13 embedded PostScript figure

Dual action antifungal small molecule modulates multidrug efflux and TOR signaling.

There is an urgent need for new strategies to treat invasive fungal infections, which are a leading cause of human mortality. Here, we establish two activities of the natural product beauvericin, which potentiates the activity of the most widely deployed class of antifungal against the leading human fungal pathogens, blocks the emergence of drug resistance, and renders antifungal-resistant pathogens responsive to treatment in mammalian infection models. Harnessing genome sequencing of beauvericin-resistant mutants, affinity purification of a biotinylated beauvericin analog, and biochemical and genetic assays reveals that beauvericin blocks multidrug efflux and inhibits the global regulator TORC1 kinase, thereby activating the protein kinase CK2 and inhibiting the molecular chaperone Hsp90. Substitutions in the multidrug transporter Pdr5 that enable beauvericin efflux impair antifungal efflux, thereby impeding resistance to the drug combination. Thus, dual targeting of multidrug efflux and TOR signaling provides a powerful, broadly effective therapeutic strategy for treating fungal infectious disease that evades resistance

Hysteretic behavior of the vortex lattice at the onset of the second peak for HgBa2_2CuO4+δ_{4+\delta} superconductor

By means of local Hall probe ac and dc permeability measurements we investigated the phase diagram of vortex matter for the HgBa$_2$CuO$_{4+\delta }$ superconductor in the regime near the critical temperature. The second peak line, $H_{\rm sp}$, in contrast to what is usually assumed, doesn't terminate at the critical temperature. Our local ac permeability measurements revealed pronounced hysteretic behavior and thermomagnetic history effects near the onset of the second peak, giving evidence for a phase transition of vortex matter from an ordered qausilattice state to a disordered glass

Phase Behavior of Type-II Superconductors with Quenched Point Pinning Disorder: A Phenomenological Proposal

A general phenomenology for phase behaviour in the mixed phase of type-II superconductors with weak point pinning disorder is outlined. We propose that the ``Bragg glass'' phase generically transforms via two separate thermodynamic phase transitions into a disordered liquid on increasing the temperature. The first transition is into a glassy phase, topologically disordered at the largest length scales; current evidence suggests that it lacks the long-ranged phase correlations expected of a ``vortex glass''. This phase has a significant degree of short-ranged translational order, unlike the disordered liquid, but no quasi-long range order, in contrast to the Bragg glass. This glassy phase, which we call a ``multi-domain glass'', is confined to a narrow sliver at intermediate fields, but broadens out both for much larger and much smaller field values. The multi-domain glass may be a ``hexatic glass''; alternatively, its glassy properties may originate in the replica symmetry breaking envisaged in recent theories of the structural glass transition. Estimates for translational correlation lengths in the multi-domain glass indicate that they can be far larger than the interline spacing for weak disorder, suggesting a plausible mechanism by which signals of a two-step transition can be obscured. Calculations of the Bragg glass-multi-domain glass and the multi-domain glass-disordered liquid phase boundaries are presented and compared to experimental data. We argue that these proposals provide a unified picture of the available experimental data on both high-T$_c$ and low-T$_c$ materials, simulations and current theoretical understanding.Comment: 70 pages, 9 postscript figures, modified title and minor changes in published versio

History of protein-protein interactions: From egg-white to complex networks.

Today, it is widely appreciated that proteinprotein interactions play a fundamental role in biological processes. This was not always the case. The study of protein interactions started slowly and evolved considerably, together with conceptual and technological progress in different areas of research through the late 19th and the 20th centuries. In this review, we present some of the key experiments that have introduced major conceptual advances in biochemistry and molecular biology, and review technological breakthroughs that have paved the way for today's systems-wide approaches to proteinprotein interaction analysis

Thermally Unstable Cooling Stimulated by Uplift: The Spoiler Clusters

Chandra X-ray observations are analyzed for five galaxy clusters whose atmospheric cooling times, entropy parameters, and ratios of cooling time to freefall time within the central galaxies lie below 1 Gyr, below 30 keV cm2, and between 20 lesssim min(t cool/t ff) lesssim 50, respectively. These thermodynamic properties are commonly associated with molecular clouds, bright Hα emission, and star formation in central galaxies. However, all have Hα luminosities below 1040 erg s−1 in the ACCEPT database. Star formation and molecular gas are absent at the levels seen in other central galaxies with similar atmospheric properties. Only RBS 0533 may host a radio/X-ray bubble, which are commonly observed in cooling atmospheres. Signatures of uplifted, high-metallicity atmospheric gas are absent. Their atmospheres are apparently thermodynamically stable despite the absence of strong nuclear feedback. We suggest that extended filaments of nebular emission and associate molecular clouds are absent at appreciable levels because their central radio sources have failed to lift low-entropy atmospheric gas to an altitude where the ratio of the cooling time to the freefall time falls below unity and the gas becomes thermally unstable

Isolation of protein complexes involved in mitosis and cytokinesis from Drosophila cultured cells

The identification of all the individual components that constitute the plethora of complexes in each cell type represents perhaps the most exciting challenge of postgenomic biology. This is particularly important in the study of events such as mitosis and cytokinesis, in which rapid and precise protein-protein interactions regulate both the direction and accuracy of these intricate processes. Here we describe an experimental strategy to isolate protein complexes involved in mitosis and cytokinesis in cultured Drosophila cells. This method involves the tagging of the bait protein with two IgG binding domains of Protein A and the isolation of the tagged bait along with its interacting partners by a single affinity purification step. These isolated complexes can then be analysed by several methods including mass spectrometry and Western blotting. Although this method has proven very successful in isolating mitotic and cytokinetic complexes, it can also be used to characterise protein complexes involved in many other cellular processes