Interaction of rat liver lysosomal membranes with actin.

Membranes were prepared from lysosomes purified 80-fold by centrifugation in a discontinuous metrizamide gradient. When salt-washed membranes were combined with rabbit muscle actin, an increase in viscosity could be measured using a falling ball viscometer. The lysosomal membrane-actin interaction was actin- and membrane-concentration dependent and appeared to be optimal under presumed physiological conditions (2 mM MgCl2, 1 mM MgATP, neutral pH, and free calcium concentration less than 10(-8) M). The actin cross-linking activity of the membrane was optimal at pH 6.4. The interaction was maximal between 10(-7) and 10(-9) M free calcium ions and inhibited by approximately 50% at concentrations of calcium greater than 0.5 x 10(-7) M. The actin-lysosomal membrane interaction was destroyed if the membranes were pretreated with Pronase, or if the membranes were purified in the absence of protease inhibitors. The interaction was not destroyed if membranes were washed with high salt or extracted with KCl and urea. In addition, a sedimentation assay for the actin-lysosomal membrane interaction was also performed to corroborate the viscometry data. The results suggest the existence of an integral lysosomal membrane actin-binding protein

Interaction of the plasma tail of comet Bradfield 1979L on 1980 February 6 with a possibly flare-generated solar-wind disturbance

Solar wind plasma data from the ISEE-3 and Helios 2 spacecraft were examined to explain a uniquely rapid 10 deg turning of the plasma tail of comet Bradfield 1979L on 1980 February 6. It was suggested that the tail position angle change occurred in response to a solar wind velocity shear across which the polar component changed by approx. 50 km s-1. The present activity was caused by noncorotating, disturbed plasma flows probably associated with an Importance 1B solar flare

Subtle temperature-induced changes in small molecule conformer dynamics-observed and quantified by NOE spectroscopy

NOE-distance relationships are shown to be sufficiently accurate to monitor very small changes in conformer populations in response to temperature (<0.5%/10 degrees C) - in good agreement with Boltzmann-predictions, illustrating the effectiveness of accurate NOE-distance measurements in obtaining high quality dynamics as well as structural information for small molecules

Expansion of magnetic clouds in the outer heliosphere

A large amount of magnetized plasma is frequently ejected from the Sun as coronal mass ejections (CMEs). Some of these ejections are detected in the solar wind as magnetic clouds (MCs) that have flux rope signatures. Magnetic clouds are structures that typically expand in the inner heliosphere. We derive the expansion properties of MCs in the outer heliosphere from one to five astronomical units to compare them with those in the inner heliosphere. We analyze MCs observed by the Ulysses spacecraft using insitu magnetic field and plasma measurements. The MC boundaries are defined in the MC frame after defining the MC axis with a minimum variance method applied only to the flux rope structure. As in the inner heliosphere, a large fraction of the velocity profile within MCs is close to a linear function of time. This is indicative of} a self-similar expansion and a MC size that locally follows a power-law of the solar distance with an exponent called zeta. We derive the value of zeta from the insitu velocity data. We analyze separately the non-perturbed MCs (cases showing a linear velocity profile almost for the full event), and perturbed MCs (cases showing a strongly distorted velocity profile). We find that non-perturbed MCs expand with a similar non-dimensional expansion rate (zeta=1.05+-0.34), i.e. slightly faster than at the solar distance and in the inner heliosphere (zeta=0.91+-0.23). The subset of perturbed MCs expands, as in the inner heliosphere, at a significantly lower rate and with a larger dispersion (zeta=0.28+-0.52) as expected from the temporal evolution found in numerical simulations. This local measure of the expansion also agrees with the distribution with distance of MC size,mean magnetic field, and plasma parameters. The MCs interacting with a strong field region, e.g. another MC, have the most variable expansion rate (ranging from compression to over-expansion)

Multi-Mission Radioisotope Thermoelectric Generator Heat Exchangers for the Mars Science Laboratory Rover

The addition of the Multi-Mission Radioisotope Thermoelectric Generator (MMRTG) to the Mars Science Laboratory (MSL) Rover requires an advanced thermal control system that is able to both recover and reject the waste heat from the MMRTG as needed in order to maintain the onboard electronics at benign temperatures despite the extreme and widely varying environmental conditions experienced both on the way to Mars and on the Martian surface. Based on the previously successful Mars landed mission thermal control schemes, a mechanically pumped fluid loop (MPFL) architecture was selected as the most robust and efficient means for meeting the MSL thermal requirements. The MSL heat recovery and rejection system (HRS) is comprised of two Freon (CFC-11) MPFLs that interact closely with one another to provide comprehensive thermal management throughout all mission phases. The first loop, called the Rover HRS (RHRS), consists of a set of pumps, thermal control valves, and heat exchangers (HXs) that enables the transport of heat from the MMRTG to the rover electronics during cold conditions or from the electronics straight to the environment for immediate heat rejection during warm conditions. The second loop, called the Cruise HRS (CHRS), is thermally coupled to the RHRS during the cruise to Mars, and provides a means for dissipating the waste heat more directly from the MMRTG as well as from both the cruise stage and rover avionics by promoting circulation to the cruise stage radiators. A multifunctional structure was developed that is capable of both collecting waste heat from the MMRTG and rejecting the waste heat to the surrounding environment. It consists of a pair of honeycomb core sandwich panels with HRS tubes bonded to both sides. Two similar HX assemblies were designed to surround the MMRTG on the aft end of the rover. Heat acquisition is accomplished on the interior (MMRTG facing) surface of each HX while heat rejection is accomplished on the exterior surface of each HX. Since these two surfaces need to be at very different temperatures in order for the fluid loops to perform efficiently, they need to be thermally isolated from one another. The HXs were therefore designed for high in-plane thermal conductivity and extremely low through-thickness thermal conductivity by using aluminum facesheets and aerogel as insulation inside a composite honeycomb core. Complex assemblies of hand-welded and uniquely bent aluminum tubes are bonded onto each side of the HX panels, and are specifically designed to be easily mated and demated to the rest of the RHRS in order to ease the integration effort

Improved NOE fitting for flexible molecules based on molecular mechanics data – a case study with S-adenosylmethionine

The use of molecular dynamics (MD) calculations to derive relative populations of conformers is highly sensitive to both timescale and parameterisation of the MD. Where these calculations are coupled with NOE data to determine the dynamics of a molecular system, this can present issues if these populations are thus relied upon. We present an approach that refines the highly accurate PANIC NMR methodology combined with clustering approaches to generate conformers, but without restraining the simulations or considering the relative population distributions generated by MD. Combining this structural sampling with NOE fitting, we demonstrate, for S-adenosylmethionine (aqueous solution at pH 7.0), significant improvements are made to the fit of populations to the experimental data, revealing a strong overall preference for the syn conformation of the adenosyl group relative to the ribose ring, but with less discrimination for the conformation of the ribose ring itself

Characterization of the Biosynthesis, Processing and Kinetic Mechanism of Action of the Enzyme Deficient in Mucopolysaccharidosis IIIC

Heparin acetyl-CoA:alpha-glucosaminide N-acetyltransferase (N-acetyltransferase, EC 2.3.1.78) is an integral lysosomal membrane protein containing 11 transmembrane domains, encoded by the HGSNAT gene. Deficiencies of N-acetyltransferase lead to mucopolysaccharidosis IIIC. We demonstrate that contrary to a previous report, the N-acetyltransferase signal peptide is co-translationally cleaved and that this event is required for its intracellular transport to the lysosome. While we confirm that the N-acetyltransferase precursor polypeptide is processed in the lysosome into a small amino-terminal alpha- and a larger ß- chain, we further characterize this event by identifying the mature amino-terminus of each chain. We also demonstrate this processing step(s) is not, as previously reported, needed to produce a functional transferase, i.e., the precursor is active. We next optimize the biochemical assay procedure so that it remains linear as N-acetyltransferase is purified or protein-extracts containing N-acetyltransferase are diluted, by the inclusion of negatively charged lipids. We then use this assay to demonstrate that the purified single N-acetyltransferase protein is both necessary and sufficient to express transferase activity, and that N-acetyltransferase functions as a monomer. Finally, the kinetic mechanism of action of purified N-acetyltransferase was evaluated and found to be a random sequential mechanism involving the formation of a ternary complex with its two substrates; i.e., N-acetyltransferase does not operate through a ping-pong mechanism as previously reported. We confirm this conclusion by demonstrating experimentally that no acetylated enzyme intermediate is formed during the reaction

Unusual composition of the solar wind in the 2–3 May 1998 CME observed with SWICS on ACE

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/95193/1/grl11810.pd

Investigation of laminar shocks

Studies of laminar shocks based on ISEE observations of terrestrial bow shocks, simulation, and theory are presented