Methicillin‐resistant Staphylococcus aureus (MRSA) poses a major threat to human health, particularly through hospital acquired infection. The spread of MRSA means that novel targets are required to ...develop potential inhibitors to combat infections caused by such drug‐resistant bacteria. Thymidylate kinase (TMK) is attractive as an antibacterial target as it is essential for providing components for DNA synthesis. Here, we report crystal structures of unliganded and thymidylate‐bound forms of S. aureus thymidylate kinase (SaTMK). His‐tagged and untagged SaTMK crystallize with differing lattice packing and show variations in conformational states for unliganded and thymidylate (TMP) bound forms. In addition to open and closed forms of SaTMK, an intermediate conformation in TMP binding is observed, in which the site is partially closed. Analysis of these structures indicates a sequence of events upon TMP binding, with helix α3 shifting position initially, followed by movement of α2 to close the substrate site. In addition, we observe significant conformational differences in the TMP‐binding site in SaTMK as compared to available TMK structures from other bacterial species, Escherichia coli and Mycobacterium tuberculosis as well as human TMK. In SaTMK, Arg 48 is situated at the base of the TMP‐binding site, close to the thymine ring, whereas a cis‐proline occupies the equivalent position in other TMKs. The observed TMK structural differences mean that design of compounds highly specific for the S. aureus enzyme looks possible; such inhibitors could minimize the transfer of drug resistance between different bacterial species.
The X-ray crystal structure of
Mycobacterium tuberculosis shikimate kinase (SK) with bound shikimate and adenosine diphosphate (ADP) has been determined to a resolution of 2.15 Å. The binding of ...shikimate in a shikimate kinase crystal structure has not previously been reported. The substrate binds in a pocket lined with hydrophobic residues and interacts with several highly conserved charged residues including Asp34, Arg58, Glu61 and Arg136 which project into the cavity. Comparisons of our ternary SK–ADP–shikimate complex with an earlier binary SK–ADP complex show that conformational changes occur on shikimate binding with the substrate-binding domain rotating by 10°. Detailed knowledge of shikimate binding is an important step in the design of inhibitors of SK, which have potential as novel anti-tuberculosis agents.
The Society for Academic Continuing Medical Education commissioned a study to clarify and, if possible, to standardize the terminology for a set of important educational interventions. In the form of ...a guideline, this article describes one such intervention, interprofessional education (IPE), which is a common intervention in health professions education. IPE is an opportunity for individuals of multiple professions to interact to learn together, to break down professional silos, and to achieve interprofessional learning outcomes in the service of high-value patient care. Based on a review of recent evidence and a facilitated discussion with US and Canadian experts, we describe IPE, its terminology, and other important information about the intervention. We encourage leaders and researchers to consider and to build on this guideline as they plan, implement, evaluate, and report IPE efforts. Clear and consistent use of terminology is imperative, along with complete and accurate descriptions of interventions, to improve the use and study of IPE.
Plasmodium falciparum, cloning of malarial genes coding for high molecular weight antigens, isolation of fragments of genes coding for proteins of 145,000 molecular weight
Yes
Whole-systems approaches are being adopted to tackle physical inactivity. The mechanisms contributing to changes resulting from whole-systems approaches are not fully understood. The voices of ...children and families that these approaches are designed for need to be heard to understand what is working, for whom, where and in what context. This paper describes the protocol for the children and families' citizen science evaluation of the Join Us: Move, Play (JU:MP) programme, a whole-systems approach to increasing physical activity in children and young people aged 5-14 years in Bradford, UK.;
The evaluation aims to understand the lived experiences of children and families' relationship with physical activity and participation in the JU:MP programme. The study takes a collaborative and contributory citizen science approach, including focus groups, parent-child dyad interviews and participatory research. Feedback and data will guide changes within this study and the JU:MP programme. We also aim to examine participant experience of citizen science and the suitability of a citizen science approach to evaluate a whole-systems approach. Data will be analysed using framework approach alongside iterative analysis with and by citizen scientists in the collaborative citizen science study.;
Ethical approval has been granted by the University of Bradford: study one (E891-focus groups as part of the control trial, E982-parent-child dyad interviews) and study two (E992). Results will be published in peer-reviewed journals and summaries will be provided to the participants, through schools or directly. The citizen scientists will provide input to create further dissemination opportunities.
The structures of the major merozoite surface antigen of Plasmodium falciparum and the gene encoding it were indistinguishable for the Wellcome strain and the Thai clone T9/94 but different for ...clones T9/96, T9/98, and T9/101. The central portion of the gene is subject to the greatest variation in structure. The protein from all five lines was found to be posttranslationally modified by covalent addition of both carbohydrate and fatty acid.
Serum from mice hyperimmune to Plasmodium yoelii was used to screen a P. yoelii genomic DNA library. Antibodies selected from hyperimmune serum by lambda gt11 clone J7 or raised against a specific ...fusion protein or peptide produced a punctate pattern of immunofluorescence on fixed smears of parasitised erythrocytes and immunoprecipitated a 235-kDa protein apparently identical to a rhoptry protein previously implicated in red cell invasion. The cloned DNA hybridised to at least seven RsaI fragments of P. yoelii genomic DNA and to three DraI fragments of similar but not identical sequence. These results suggest that the gene encoding the 235-kDa rhoptry protein may be represented more than once in the P. yoelii genome.
We investigated the ability of retinoic acid (RA) to alter the binding of epidermal growth factor (EGF) to Rat-1 and Swiss mouse 3T3 cells, the EGF-related induction of ornithine decarboxylase (ODC) ...activity and DNA synthesis. Pretreatment of 3T3 cells with 10(-10)--10(-5) M RA produced a dose-dependent increase in EGF binding. The increased EGF binding appeared to result from an increased number of EGF receptors and did not occur in Rat-1 cells which, unlike 3T3 cells, possessed measureable amounts of cellular retinoic acid binding protein (CRABP). Both cell lines responded to RA pretreatment with an increase in EGF- and TPA-induced ODC activity, without a concomitant increase in DNA synthesis in either cell line. We suggest that the increase in EGF binding induced by RA in 3T3 cells is apparently unrelated to alterations in EGF-induced biological responses and to the presence of CRABP.
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