Real-Time Measurement of Myosin–Nucleotide Noncovalent Complexes by Electrospray Ionization Mass Spectrometry

AbstractNanoelectrospray ionization mass spectrometry has been used to measure the binding of ATP and ADP to the active site of rabbit skeletal myosin-S1. Increases in the molecular mass of myosin-S1 of 425±10Da were obtained with the binding of ADP to the active site and by 530±10Da with either ATP or hydrolysis products ADP and phosphate. Active site titrations of myosin-S1 with ADP gave a stoichiometry of ∼1 ADP/S1 with an affinity in the micromolar range. The binding of ATP to myosin-S1 could be observed in the presence of up to 60μM of excess MgATP without nonspecific binding of MgATP to the myosin. Conversion of the nucleotide complex containing an equilibrium mixture of ATP and ADP-Pi bound to myosin-S1 to one containing only bound ADP occurs at a rate consistent with that of the known steady-state rate of ATP hydrolysis. We expect this method to be of considerable use in the analysis of ligand binding and hydrolysis by the active sites of expressed myosin and myosin subfragments, which are not available in sufficient quantities for conventional methods of measurement of ligand binding

The effect of farming system on dairy cow cleanliness in the UK and implications to udder health

The cleanliness of dairy cows was assessed using a 20 point hygiene score system at different times in the year on 14 organic and 14 conventional farms in the UK. Overall, cows were dirtier during winter housing compared to summer grazing. Farming system had no effect on cow cleanliness when cows were at grass, but when housed in the winter, organic cows were more likely to be cleaner. There was a link between cow hygiene scores and milk hygiene, with herds having lower bulk tank somatic cell counts (BTSCC) tending to have cleaner cows. This relationship was strongest for the organic herds. There was no significant link between hygiene score and Bactoscan (BS) count or mastitis incidence

MePrints: Building User Centred Repositories

Over the last few years we have been working to reinvent Teaching and Learning Repositories learning from the best practices of Web 2.0. Over this time we have successfully deployed a number of innovative repositories, including Southampton University EdShare, The Language Box, The HumBox, Open University’s LORO and Worcester Learning Box

An associative information visualizer

IEEE Symposium on Information Visualization, INFOVIS 2004, p. r8

Landform Evolution Modeling of Sedimentary Processes on Icy Worlds: the Cases of Hyperion and Helene

Sedimentary activity (erosion, transportation and deposition) is a major landscape-shaping process on many icy worlds of the outer solar system for which an understanding of this process is central to their geologic characterizations. Several recent studies, lead by the authors, have identified the effects of sedimentary activity on icy satellite landforms and recognized the role of mass wasting and volatile loss and redistribution in the evolution of these features. We apply state of the art, physics based, landform evolution modeling to icy satellite sedimentary landforms in order to fully quantitatively characterize how their morphologies change with time, and what their present appearances imply for the initial abundance and distribution of loose material, volatiles, and refractories in original pristine landforms. We will present results of our landform evolution modeling to the development of the landscapes of the Saturnian moons of Hyperion and Helene. Our current conclusions for Hyperion are that its unique appearance can be explained in part by the loss to space of ballistic ejecta during impact events. In order to create the smooth surfaces and the reticulate, honeycomb pattern of narrow divides between old craters, appreciable subsequent modification of crater morphology must occur through mass-wasting processes accompanied by sublimation, probably facilitated by the loss of CO2 as a component of the relief-supporting matrix of the bedrock. This mass wasting effectively destroys small craters, at least in part accounting for the paucity of sub-kilometer craters on Hyperion. Helenes unusual morphology consists of broad depressions (modified large craters) and a generally smooth surface patterned with streaks and grooves. The streaks appear to be oriented down-gradient, as are the grooves. This pattern suggests intensive mass-wasting as a dominant process. Our initial modeling of this surface suggests a Bingham-like rheological behavior for the loose down-slope moving material. Interestingly, as a Bingham flow, the models indicate that aperiodic "intermittent" behavior to be present, suggesting that periods of quiet steady landform evolution are punctuated short periods of active surface readjustment