Calcium is essential for the structural integrity of the cysteine-rich, ligand-binding repeat of the low-density lipoprotein receptor

Seven cysteine-rich repeats form the ligand-binding region of the low-density lipoprotein (LDL) receptor. Each of these repeats is assumed to bind a calcium ion, which is needed for association of the receptor with its ligands, LDL and beta-VLDL. The effects of metal ions on the folding of the reduced N-terminal cysteine-rich repeat have been examined by using reverse-phase high-performance liquid chromatography to follow the formation of fully oxidized isomers with different disulfide connectivities. in the absence of calcium many of the 15 possible isomers formed on oxidation, whereas in its presence the predominant product at equilibrium had the native disulfide bond connectivities. Other metals were far less effective at directing disulfide bond formation: Mn2+ partly mimicked the action of Ca2+, but Ba2+, Sr2+, and Mg2+ had little effect. This metal-ion specificity was also observed in two-dimensional H-1 NMR spectral studies: only Ca2+ induced the native three-dimensional fold. The two paramagnetic ions, Gd3+ and Mn2+, and Cd2+ did not promote adoption of a well-defined structure, and the two paramagnetic ions did not displace calcium ions. The location of calcium ion binding sites in the repeat was also explored by NMR spectroscopy. The absence of chemical shift changes for the side chain proton resonances of Asp26, Asp36, and Glu37 from pH 3.9 to 6.8 in the presence of calcium ions and their proximal location in the NMR structures implicated these side chains as calcium ligands. Deuterium exchange NMR experiments also revealed a network of hydrogen bonds that stabilizes the putative calcium-binding loop.

Systems for assessing the effectiveness of management in protected areas

Since the mid-1990s, numerous methodologies have been developed to assess the management effectiveness of protected areas, many tailored to particular regions or habitats. Recognizing the need for a generic approach, the World Commission on Protected Areas (WCPA) developed an evaluation framework allowing specific evaluation methodologies to be designed within a consistent overall approach. Twenty-seven assessment methodologies were analyzed in relation to this framework. Two types of data were identified: quantitative data derived from monitoring and qualitative data derived from scoring by managers and stakeholders. The distinction between methodologies based on data types reflects different approaches to assessing management. Few methodologies assess all the WCPA framework elements. More useful information for adaptive management will come from addressing all six elements. The framework can be used to adapt existing methodologies or to design new, more comprehensive methodologies for evaluation, using quantitative monitoring data, qualitative scoring data, or a combination of both.

Kinetic and structural evidence for the importance of Tyr236 for the integrity of the Mo active site in a bacterial sulfite dehydrogenase

The sulfite dehydrogenase from Starkeya novella is the only known sulfite-oxidizing enzyme that forms a permanent heterodimeric complex between a molybdenum and a heme c-containing subunit and can be crystallized in an electron transfer competent conformation. Tyr236 is a highly conserved active site residue in sulfite oxidoreductases and has been shown to interact with a nearby arginine and a molybdenum-oxo ligand that is involved in catalysis. We have created a Tyr236 to Phe substitution in the SorAB sulfite dehydrogenase. The purified SDHY236F protein has been characterized in terms of activity, structure, intramolecular electron transfer, and EPR properties. The substituted protein exhibited reduced turnover rates and substrate affinity as well as an altered reactivity toward molecular oxygen as an electron acceptor. Following reduction by sulfite and unlike SDHWT, the substituted enzyme was reoxidized quickly in the presence of molecular oxygen, a process reminiscent of the reactions of the sulfite oxidases. SDHY236F also exhibited the pH-dependent CW-EPR signals that are typically observed in vertebrate sulfite oxidases, allowing a direct link of CW-EPR properties to changes caused by a single-amino acid substitution. No quantifiable electron transfer was seen in laser flash photolysis experiments with SDHY236F. The crystal structure of SDHY236F clearly shows that as a result of the substitution the hydrogen bonding network surrounding the active site is disturbed, resulting in an increased mobility of the nearby arginine. These disruptions underline the importance of Tyr236 for the integrity of the substrate binding site and the optimal alignment of Arg55, which appears to be necessary for efficient electron transfer.

Philosophy of Management

All seven papers in this issue on Fakes, Copies and Originals consider matters of reproduction and origination. They consider the interplay between culture meanings and financial capital, and share in common a concern for further developing the cultural wealth of society.

The role of management incentives in successful information systems development and implementation

Because organizations are making large investments in Information systems (IS), efficient IS project management has been found critical to success. This study examines how the use of incentives can improve the project success. Agency theory is used to: identify motivational factors of project success, help the IS owners to understand to what extent management incentives can improve IS development and implementation (ISD/I). The outcomes will help practitioners and researchers to build on theoretical model of project management elements which lead to project success. Given the principal-agent nature of most significant scale of IS development, insights that will allow for greater alignment of the agent’s goals with those of the principal through incentive contracts, will serve to make ISD/I both more efficient and more effective, leading to more successful IS projects.