Introduction of a stabilizing 10 residue {beta}-hairpin in Bacillus subtilis neutral protease

A 10 residue ß-hairpin, which is characteristic ofthermostable Bacillus neutral proteases, was engineered intothe thermolabile neutral protease of Bacillus subtilis. Therecipient enzyme remained fully active after introduction ofthe loop. However, the mutant protein exhibited autocatalyticnicking and a 0.4°C decrease in thermostability. Two additionalpoint mutations designed to improve the interactions betweenthe enzyme surface and the introduced ß-hairpin resultedin reduced nicking and increased thermostability. After theintroduction of both additional mutations in the loopcontainingmutant, nicking was largely prevented and an increase in thermostabilityof 1.1°C was achieved.     

The effect of cavity-filling mutations on the thermostability of Bacillus stearothermophilus neutral protease

Cavities in the hydrophobic core of the neutral protease ofBacillus stearothermophilus were analyzed using a threedimensionalmodel that was inferred from the crystal structure of thermolysin,the highly homologous neutral protease of B.thermoproteolyticus(85% sequence identity). Site–directed mutagenesis wasused to fill some of these cavities, thereby improving hydrophobicpacking in the protein interior. The mutations had small effectson the thermostability, even after drastic changes, such asLeu284Trp and Met168Trp. The effects on T50, the temperatureat which 50% of the enzyme is irreversibly inactivated in 30min, ranged from 0.0 to +0.4°C. These results can be explainedby assuming that the mutations have positive and negative structuraleffects of approximately the same magnitude. Alternatively,it could be envisaged that the local unfolding steps, whichrender the enzyme susceptible towards autolysis and which arerate limiting in the process of thermal inactivation, are onlyslightly affected by alterations in the hydrophobic core.     

Compared imbibitions of ordinary and high performance concrete with null or positive water pressure head

A method to simultaneously measure the moisture diffusion coefficient, D_θ, of unsaturated concrete, and the saturated concrete hydraulic conductivity, K_l, was developed for cylindrical specimens placed on a container filled with water that could be maintained at a given hydraulic pressure. Ordinary Portland cement Concrete (OPC) with a moderate and High Performance Concrete (HPC) with a low water to cement ratio were tested. The time dependent distribution of water content in the specimens was measured using a non-intrusive method based on gamma-ray attenuation. The measurements were conducted with varying hydraulic head (positive or null). Boltzmann's transformation was used to analyze the experimental results obtained at different hydraulic pressures and the difference between the null (or atmospheric) and positive pressure results is used to accurately determine K_l and also D_θ . This paper will present the results obtained using this original method, possible interpretations and future research.     

Elucidating a biological role for chromium at a molecular level

Chromium is an essential trace element for mammals and is required for maintenance of proper carbohydrate and lipid metabolism. However, elucidating its function at a molecular level has proved to be problematic. Recent research has revealed that the chromium-binding oligopeptide chromodulin may play a unique role in the autoamplification of insulin signaling. Attempts to develop chromium-containing nutritional supplements and therapeutics are described.     

Conformational flexibility of domain III of annexin V studied by fluorescence of tryptophan 187 and circular dichroism: the effect of pH

The conformation and dynamics of domain III of annexin V was studied by steady-state and time-resolved fluorescence of its single tryptophan residue (Trp187) as a function of pH in the absence of calcium. At neutral pH, the maximum of emission occurs at 326 nm, in agreement with the hydrophobic location of the tryptophan residue seen in the three-dimensional structure. Upon decreasing the pH, a progressive red-shift by about 12 nm of the fluorescence emission spectrum is observed. The effect is complete between pH 6 and 4.5, and most likely involves at least one and maybe two carboxylic group(s). Circular dichroism mesurements give evidence for a preservation of the native folding of the protein in these mild acidic conditions. A fluorescence red-shift of smaller amplitude is also observed at high pH (approximately 11). The aggregation state of the protein is affected by pH: while at neutral pH, the protein is monomeric (rotational correlation time = 14 ns); it forms aggregates larger than a dimer (rotational correlation time > 40 ns) in acidic pH conditions. These results suggest that electrostatic interactions are probably important for the stabilization of the folding of domain III without calcium. The conformational change may be related to the aggregation state of the molecule. Examination of the protein crystal structures with and without calcium ion in domain III shows an interplay of salt bridges implying charged amino acid side chains at the molecule surface of domain III. These observations may provide a further clue to the mechanism of the conformational change of domain III of annexin V induced by high calcium concentrations and interaction at the membrane/water interface.     

An Ordering Model for Recoverable Stock Items

A recoverable stock item is one which after use is repaired and used again. This article presents a minimum investment model for ordering new stock when the units available through repair do not satisfy total demand. Simulation is utilized to measure the effect of the required approximations.     

An Overview of Progress in the Design And Implementation of Landsat-D Systems

Landsat-D will be launched in late 1981 or early 1982 into a Sun-synchronous orbit near 700 km. Landsat-D systems and the principal observing instrument, the Thematic Mapper, are being designed and implemented to provide a significantly improved Earth-resources monitoring capability. The Thematic Mapper will have seven spectral bands (0.45-0.52, 0.52-0.60, 0.63-0.69, 0.76-0.90, 1.55-1.75, 2.08-2.35, and 10.5-12.5 ?m), with an instantaneous field of view at nadir of 30 m except for the thermal band (120 m). The data from the Thematic Mapper and other Landsat-D flight segment systems will be relayed to the Landsat-D ground data processing system using a direct readout capability or the Tracking and Data Relay Satellite System and communications satellites. The ground systems will process the data as it arrives at rates up to 100 Mbit/s and a total volume of 2.6 × 1011 bit/day into standard digital and photographic products within 48 hours of data acquisition. This includes 200 Multispectral Scanner Subsystem and 100 Thematic Mapper scenes per day. The Thematic Mapper increased spatial, spectral, and radiometric resolution should be of primary benefit in agricultural and vegetation surveys, but substantial improvements in resource and land cover observations for geology, land use, and water resources applications are also to be anticipated.     

Composition and transfer mechanism of Langmuir-Blodgett multilayers of stearates

We determined the composition of Langmuir-Blodgett multilayers of several stearates from various experimental data and related these results to the type (Y or XY) of the layers. It is shown that the transfer of the XY layers can be accounted for without invoking an overturning mechanism.     

The extended jones—dole equation

A number of authors have recently interpreted the relative viscosities of electrolytic solutions in terms of an extended Jones—Dole equation containing a term Dc²:

The B-coefficients resulting from such analyses are not directly comparable with those arising from the simple Jones—Dole expression because of the high negative correlation between the B and D terms. There is therefore some difficulty in relating results from both types of analysis. In this paper a number of data sets are examined statistically with a view to establishing a standard procedure for obtaining estimates of the B-coefficients which relate exclusively to the linear term and are not influenced by the effect of higher terms. Two methods which recognize the high correlation of B and D terms are suggested.