Structural Studies of a Stabilized Phosphoenzyme Intermediate of Ca 2+ -ATPase

Ca(2+)-ATPase belongs to the family of P-type ATPases and maintains low concentrations of intracellular Ca(2+). Its reaction cycle consists of four main intermediates that alternate ion binding in the transmembrane domain with phosphorylation of an aspartate residue in a cytoplasmic domain. Previous work characterized an ultrastable phosphoenzyme produced first by labeling with fluorescein isothiocyanate, then by allowing this labeled enzyme to establish a maximal Ca(2+) gradient, and finally by removing Ca(2+) from the solution. This phosphoenzyme is characterized by very low fluorescence and has specific enzymatic properties suggesting the existence of a high energy phosphoryl bond. To study the structural properties of this phosphoenzyme, we used cryoelectron microscopy of two-dimensional crystals formed in the presence of decavanadate and determined the structure at 8-A resolution. To our surprise we found that at this resolution the low fluorescence phosphoenzyme had a structure similar to that of the native enzyme crystallized under equivalent conditions. We went on to use glutaraldehyde cross-linking and proteolysis for independent structural assessment and concluded that, like the unphosphorylated native enzyme, Ca(2+) and vanadate exert a strong influence over the global structure of this low fluorescence phosphoenzyme. Based on a structural model with fluorescein isothiocyanate bound at the ATP site, we suggest that the stability as well as the low fluorescence of this phosphoenzyme is due to a fluorescein-mediated cross-link between two cytoplasmic domains that prevents hydrolysis of the aspartyl phosphate. Finally, we consider the alternative possibility that phosphate transfer to fluorescein itself could explain the properties of this low fluorescence species

The Deposit Policy

The chapter version presented here is the authors' authorized manuscript. Pagination per the published monograph is indicated in this manuscript with the page numbers near the beginning or ending words per the publication, highlighted in green for the start of the page and red for the end.There are a host of implications and risks associated with the act of placing any author’s writing in the public sphere. As in the case of an institutional repository (IR), the mere fact of a university declaring: “Here are works related to this institution” is a bold act that opens up vulnerabilities just as it allows that organization to establish its identity, demonstrate its importance and quality, and publicize its impactful contributions to society. What is a university to do

Deicing Composition

A deicing composition is provided. The composition includes a potassium or sodium salt of a carboxylic acid and a lithium salt of a carboxylic acid or lithium nitrate, wherein the molar ratio of lithium to potassium or lithium to sodium is from 10 percent to 80 percent

Deicing composition

A deicing composition is provided. The composition includes a potassium or sodium salt of a carboxylic acid and a lithium salt of a carboxylic acid or lithium nitrate, wherein the molar ratio of lithium to potassium or lithium to sodium is from 10 percent to 80 percent

Report of the DAMENames Ad Hoc Committee

In early 2018, the DAMEid group requested that Cataloging and Metadata unit examine the metadata needs for the DAME. When analyzing metadata needs in both OAKTrust and Fedora, it became clear that the lack of name authority control was causing serious problems for users, especially in the case of a single author having many entries in the author index. For example, Steven M. Wright, Royce E. Wisenbaker, Professor II in Chemical and Electrical Engineering, has 10 different entries for his name. This problem is caused by the lack of authority control and the inconsistent ways in which names are inputted into Vireo and OAKTrust. In their report to the DAMEid committee, the Metadata and Cataloging librarians strongly suggested that some type of name authority control be implemented within the DAME. In smaller repositories with few names and fewer entities (e.g., persons, organizations, subjects, etc.), the absence of explicit disambiguation or authority control can be a manageable problem. When only a few authors share a name, it is easy to tell them apart based on the subject matter of the works attached to the name. The problem compounds as collections grow larger and the number of entities with the same name that need to be distinguished from each other increases. For example, in the large OAKTrust IR, it is hard for a user to identify the "Steven Wright" that he or she is looking for, as there are several authors so named with dozens of items in the IR. Another issue that emerges in a system with no authority control – such as OAKTrust – is that an everyday typographical error (an extra space, no period after an initial, misspellings, etc.) results in a new entry in the author list. This results in multiple names for one person and it means that there is no way for a user to easily identify all the works attributed to one author

Climate Clever Beef : On-farm demonstration of adaptation and mitigation options for climate change in northern Australia

This project engaged with beef producers in five regions of northern Australia to identify management options that improve the performance and resilience of beef businesses. The work was done in the context of increasing the resilience of businesses to current climate variability as well as to projected changes in climate. The project also identified potential synergies and conflicts between improved business performance, climate adaptation practices and greenhouse gas emissions management. Three regions (Qld Gulf, Fitzroy Basin and Victoria River District) evaluated climate adaptation and greenhouse gas mitigation options via a benchmarking and options analysis approach with three “focal” properties. Five indicators of performance were evaluated for each property – profitability, productivity, land condition, climate change risk and greenhouse gas emissions. These detailed business analyses were complemented by demonstration sites in each region. Two other regions (Qld Mitchell grasslands and NT Barkly Tablelands) used on-property demonstration sites to showcase promising climate adaptation practices identified in a previous project. The focal property approach provided a systematic process for assessing current business performance as well as a ready means of estimating the impacts of management changes. For example, over a 15 year period, the Qld Gulf focal property improved its pasture condition dramatically by stocking around the long-term carrying capacity and undertaking wet season spelling. This, combined with herd management improvements, increased profitability and productivity, reduced greenhouse gas emissions by 15%, and improved greenhouse gas emission efficiency by >100%. The demonstration sites in each region effectively promoted and documented the benefits of key grazing practices for improving resilience to both current climate variability and potential climate change. For example, on a 16,118km² property in the Barkly, a paddock demonstration has documented initial land condition recovery at old bores, and the reduced risk of overgrazing around new bores, through best practice stocking rate management and wet season spelling. Qualitative analyses showed that many of the adaptation practices identified for improving resilience are consistent with existing best practice recommendations aimed at improving productivity and sustainability. Furthermore, these adaptation practices appear to have largely neutral implications for greenhouse gas emissions. In contrast, practices and options for reducing greenhouse gas emissions were more likely to create conflicts that leave enterprises more vulnerable to climate change. Examples of the negative consequences of mitigation measures include reduced pasture production associated with increased carbon sequestration in trees (i.e. woody vegetation thickening or regrowth retention) and increased operating costs associated with carbon pricing (if these are not offset with carbon credits). The project demonstration sites and focal property benchmarking process provided a solid base for focussed extension work targeting the drivers of profit, land condition, greenhouse gas emissions intensity and climate adaptation strategies

CRUZ, MARUJA [Material gráfico]

Copia digital. Madrid : Ministerio de Educación, Cultura y Deporte. Subdirección General de Coordinación Bibliotecaria, 201