Structure, substrate recognition and reactivity of Leishmania major mevalonate kinase

This research was supported by the German Academic Exchange Service (DAAD), the Wellcome Trust (TKS and WNH as Trust Senior Research fellows), the Biotechnology and Biological Science Research Council (Structural Proteomics of Rational Targets) and the European Synchrotron Radiation Facility.Background: Isoprenoid precursor synthesis via the mevalonate route in humans and pathogenic trypanosomatids is an important metabolic pathway. There is however, only limited information available on the structure and reactivity of the component enzymes in trypanosomatids. Since isoprenoid biosynthesis is essential for trypanosomatid viability and may provide new targets for therapeutic intervention it is important to characterize the pathway components. Results: Putative mevalonate kinase encoding genes from Leishmania major (LmMK) and Trypanosoma brucei (TbMK) have been cloned, over-expressed in and proteins isolated from procyclic-form T. brucei. A highly sensitive radioactive assay was developed and shows ATP-dependent phosphorylation of mevalonate. Apo and (R)-mevalonate bound crystal structures of LmMK, from a bacterial expression system, have been determined to high resolution providing, for the first time, information concerning binding of mevalonate to an MK. The mevalonate binds in a deep cavity lined by highly conserved residues. His25 is key for binding and for discrimination of (R)-over (S)-mevalonate, with the main chain amide interacting with the C3 hydroxyl group of ( R)mevalonate, and the side chain contributing, together with Val202 and Thr283, to the construction of a hydrophobic binding site for the C3 methyl substituent. The C5 hydroxyl, where phosphorylation occurs, points towards catalytic residues, Lys18 and Asp155. The activity of LmMK was significantly reduced compared to MK from other species and we were unable to obtain ATP-binding data. Comparisons with the rat MK:ATP complex were used to investigate how this substrate might bind. In LmMK, helix alpha 2 and the preceding polypeptide adopt a conformation, not seen in related kinase structures, impeding access to the nucleotide triphosphate binding site suggesting that a conformational rearrangement is required to allow ATP binding. Conclusion: Our new structural information, consistent with data on homologous enzymes allows a detailed description of how mevalonate is recognized and positioned for catalysis in MK. The mevalonate-binding site is highly conserved yet the ATP-binding site is structurally distinct in LmMK. We are unable to provide a definitive explanation for the low activity of recombinant protein isolated from a bacterial expression system compared to material isolated from procyclic-form Trypanosoma brucei.Publisher PDFPeer reviewe

DO MISSING INSTITUTIONAL ARRANGEMENTS BLOCK ENLARGEMENT OF WATER MARKETS?

As we consider the potential for expanding water markets as a means to help prevent water shortages, it is clear that there is resistance to such an expansion. This resistance should not be surprising given that there are likely to be both gainers and losers from expanded water markets. This papers shows that there are a number of potential market failures when water markets are expanded and that these failures are important to different stakeholder groups. These failures result from both technical and pecuniary externalities. If markets are to expand beyond the local level, new institutional arrangements will be needed that help reduce the negative impacts of the different market failures. In the past a number of institutional arrangements have been used by different stakeholder groups to block trading. We review some of these arrangements as well as institutional arrangements that can work to promote market expansion by mitigating market failures or by compensating damaged parties.Resource /Energy Economics and Policy,

Temperature sensitivity of Eppley broadband radiometers

Broadband radiometers manufactured by Eppley Laboratories Inc. are commonly used to measure irradiance from both ground-based and aircraft platforms. Namely, the pyranometer (Model PSP) measures irradiance in the .3 to 3.0 micron spectral region while the pyrgeometer (Model PIR) senses energy in the 4 to 50 micron region. The two instruments have a similar thermopile construction but different filters to achieve the appropriate spectral selection. During the fall of 1986, the First ISCCP (International Satellite Cloud Climatology Project) Regional Experiment (FIRE) commenced with the first cirrus Intensive Field Observation (IFO) conducted in Central Wisconsin. Due to the nature of this field project, pyranometers and pyrgeometers manufactured by Eppley were flown on NCAR's high altitude research aircraft, the Sabreliner. Inherent in the construction of these radiometers is temperature compensation circuitry designed to make the instrument sensitivity nominally constant over a temperature range from -20 to +40 C. Because the Sabreliner flew at high altitudes where temperatures were as cold as -70 C, it was necessary to determine the radiometers relative sensitivity to temperatures below -20 C and apply appropriate corrections to the FIRE radiation data set. A procedure to perform this calibration is outlined. It is meant to serve as a supplement to calibration procedures

Radiative properties of cirrus clouds inferred from broadband measurements during FIRE

It is well known that clouds are significant modulators of weather and climate because of their effects on the radiation field and thus on the energy balance of the earth atmosphere system. As a result, the accurate prediction of weather and climate depends to a significant degree on the accuracy with which cloud radiation interactions can be described. The broadband radiative and microphysical properties of five cirrus cloud systems are reported, as observed from the NCAR Sabreliner during the FIRE first Cirrus IFO, in order to better understand cirrus cloud-radiation interactions. A broadband infrared (BBIR) radiative transfer model is used to deduce BBIR absorption coefficients in order to assess the impact of the cirrus clouds on infrared radiation. The relationships of these absorption coefficients to temperature and microphysical characteristics are explored

WATER MARKETS AND THIRD PARTY EFFECTS

This paper examines the potential effects of water trading on the service sector of a rural economy. The economy earns income producing an irrigated agricultural product and a non-agricultural (service) good, and possibly by selling water. Among other things, we show when none of the water income leaves the region (no income flight), water trading enhances regional welfare. We then show if income flight is "large enough", water trading has the opposite effect. Albeit, even under income flight, if the income flight problem is not too serious, water trading will enhance regional welfare.Resource /Energy Economics and Policy,

Water Markets and Third Party Effects

We examine the potential effects of water trading on aggregate welfare and income distribution across the agricultural and service sector of a small rural economy. We show that per capita welfare (real income) of agents in the region increase with increased water trading. Not surprisingly, if enough agents leaves the region (income flight), nominal income can fall. If the share of household income spent on services is large (small) relative to the cost share of services in agricultural production, then the service price increases (decreases) with increased water trading. Typically when the service price falls (increases) farmers win (lose), service providers lose (win) and agricultural service providers almost always lose. Thus, a natural conflict emerges between farming and service sector stakeholders.Resource /Energy Economics and Policy, Q25, Q28, R0,

Radiative properties of Cirrus clouds: FIRE IFO case October 28, 1986

A description of the radiative properties of two cirrus clouds sampled on 10/28/88 in the FIRE cirrus IFO is presented. The clouds are characterized in terms of the broadband infrared effective emittance, cloud fractional absorptance, shortwave heating rate, cloud albedo and vertical velocity. The broadband fluxes used in these calculations were obtained from measurements made by pyranometers and pyrgeometers. The shortwave irradiances were corrected to a horizontal plane and normalized to the same time by taking into account Sabreliner flight information (i.e., pitch, roll, heading and angle of attack), as well as sun-earth geometry considerations. Since only one aircraft was used, broadband fluxes at different levels in the cloud were not measured simultaneously. As a result, sampling errors may occur due to the nonsteady state of the cloud field or due to the possibility that the flight legs were not flown directly above or below each other. To minimize these errors and to simplify the analysis, the necessary variables were averaged and the averages used in the calculations. The downwelling shortwave and longwave irradiances were used as selection criteria to remove cloud free data encountered along the data sampling leg