Formal Reduction Potential of 3,5-Difluorotyrosine in a Structured Protein: Insight into Multistep Radical Transfer

The reversible Y–O•/Y–OH redox properties of the α[subscript 3]Y model protein allow access to the electrochemical and thermodynamic properties of 3,5-difluorotyrosine. The unnatural amino acid has been incorporated at position 32, the dedicated radical site in α[subscript 3]Y, by in vivo nonsense codon suppression. Incorporation of 3,5-difluorotyrosine gives rise to very minor structural changes in the protein scaffold at pH values below the apparent pK (8.0 ± 0.1) of the unnatural residue. Square-wave voltammetry on α[subscript 3](3,5)F[subscript 2]Y provides an E°′(Y–O•/Y–OH) of 1026 ± 4 mV versus the normal hydrogen electrode (pH 5.70 ± 0.02) and shows that the fluoro substitutions lower the E°′ by −30 ± 3 mV. These results illustrate the utility of combining the optimized α[subscript 3]Y tyrosine radical system with in vivo nonsense codon suppression to obtain the formal reduction potential of an unnatural aromatic residue residing within a well-structured protein. It is further observed that the protein E°′ values differ significantly from peak potentials derived from irreversible voltammograms of the corresponding aqueous species. This is notable because solution potentials have been the main thermodynamic data available for amino acid radicals. The findings in this paper are discussed relative to recent mechanistic studies of the multistep radical-transfer process in Escherichia coli ribonucleotide reductase site-specifically labeled with unnatural tyrosine residues.National Institutes of Health (U.S.) (Grant GM29595

Tropomyosin - master regulator of actin filament function in the cytoskeleton.

Tropomyosin (Tpm) isoforms are the master regulators of the functions of individual actin filaments in fungi and metazoans. Tpms are coiled-coil parallel dimers that form a head-to-tail polymer along the length of actin filaments. Yeast only has two Tpm isoforms, whereas mammals have over 40. Each cytoskeletal actin filament contains a homopolymer of Tpm homodimers, resulting in a filament of uniform Tpm composition along its length. Evidence for this ‘master regulator’ role is based on four core sets of observation. First, spatially and functionally distinct actin filaments contain different Tpm isoforms, and recent data suggest that members of the formin family of actin filament nucleators can specify which Tpm isoform is added to the growing actin filament. Second, Tpms regulate whole-organism physiology in terms of morphogenesis, cell proliferation, vesicle trafficking, biomechanics, glucose metabolism and organ size in an isoform-specific manner. Third, Tpms achieve these functional outputs by regulating the interaction of actin filaments with myosin motors and actin-binding proteins in an isoform-specific manner. Last, the assembly of complex structures, such as stress fibers and podosomes involves the collaboration of multiple types of actin filament specified by their Tpm composition. This allows the cell to specify actin filament function in time and space by simply specifying their Tpm isoform composition

Synthesis Paper: Targeted Livestock Grazing: Prescription for Healthy Rangelands

Targeted livestock grazing is a proven tool for manipulating rangeland vegetation, and current knowledge about targeted livestock grazing is extensive and expanding rapidly. Targeted grazing prescriptions optimize the timing, frequency, intensity, and selectivity of grazing (or browsing) in combinations that purposely exert grazing/browsing pressure on specific plant species or portions of the landscape. Targeted grazing differs from traditional grazing management in that the goal of targeted grazing is to apply defoliation or trampling to achieve specific vegetation management objectives, whereas the goal of traditional livestock grazing management is generally the production of livestock commodities. A shared aim of targeted livestock grazing and traditional grazing management is to sustain healthy soils, flora, fauna, and water resources that, in turn, can sustain natural ecological processes (e.g., nutrient cycle, water cycle, energy flow). Targeted grazing prescriptions integrate knowledge of plant ecology, livestock nutrition, and livestock foraging behavior. Livestock can be focused on target areas through fencing, herding, or supplement placement. Although practices can be developed to minimize the impact of toxins contained in target plants, the welfare of the animals used in targeted grazing must be a priority. Monitoring is needed to determine if targeted grazing is successful and to refine techniques to improve efficacy and efficiency. Examples of previous research studies and approaches are presented to highlight the ecological benefits that can be achieved when targeted grazing is applied properly. These cases include ways to suppress invasive plants and ways to enhance wildlife habitat and biodiversity. Future research should address the potential to select more adapted and effective livestock for targeted grazing and the associated animal welfare concerns with this practice. Targeted livestock grazing provides land managers a viable alternative to mechanical, chemical, and prescribed fire treatments to manipulate rangeland vegetation

The Potential of Antimicrobial Peptides as Biocides

Antimicrobial peptides constitute a diverse class of naturally occurring antimicrobial molecules which have activity against a wide range of pathogenic microorganisms. Antimicrobial peptides are exciting leads in the development of novel biocidal agents at a time when classical antibiotics are under intense pressure from emerging resistance, and the global industry in antibiotic research and development stagnates. This review will examine the potential of antimicrobial peptides, both natural and synthetic, as novel biocidal agents in the battle against multi-drug resistant pathogen infections

Message Journal, Issue 5: COVID-19 SPECIAL ISSUE Capturing visual insights, thoughts and reflections on 2020/21 and beyond...

If there is a theme running through the Message Covid-19 special issue, it is one of caring. Of our own and others’ resilience and wellbeing, of friendship and community, of students, practitioners and their futures, of social justice, equality and of doing the right thing. The veins of designing with care run through the edition, wide and deep. It captures, not designers as heroes, but those with humble views, exposing the need to understand a diversity of perspectives when trying to comprehend the complexity that Covid-19 continues to generate. As graphic designers, illustrators and visual communicators, contributors have created, documented, written, visualised, reflected, shared, connected and co-created, designed for good causes and re-defined what it is to be a student, an academic and a designer during the pandemic. This poignant period in time has driven us, through isolation, towards new rules of living, and new ways of working; to see and map the world in a different light. A light that is uncertain, disjointed, and constantly being redefined. This Message issue captures responses from the graphic communication design community in their raw state, to allow contributors to communicate their experiences through both their written and visual voice. Thus, the reader can discern as much from the words as the design and visualisations. Through this issue a substantial number of contributions have focused on personal reflection, isolation, fear, anxiety and wellbeing, as well as reaching out to community, making connections and collaborating. This was not surprising in a world in which connection with others has often been remote, and where ‘normal’ social structures of support and care have been broken down. We also gain insight into those who are using graphic communication design to inspire and capture new ways of teaching and learning, developing themselves as designers, educators, and activists, responding to social justice and to do good; gaining greater insight into society, government actions and conspiracy. Introduction: Victoria Squire - Coping with Covid: Community, connection and collaboration: James Alexander & Carole Evans, Meg Davies, Matthew Frame, Chae Ho Lee, Alma Hoffmann, Holly K. Kaufman-Hill, Joshua Korenblat, Warren Lehrer, Christine Lhowe, Sara Nesteruk, Cat Normoyle & Jessica Teague, Kyuha Shim. - Coping with Covid: Isolation, wellbeing and hope: Sadia Abdisalam, Tom Ayling, Jessica Barness, Megan Culliford, Stephanie Cunningham, Sofija Gvozdeva, Hedzlynn Kamaruzzaman, Merle Karp, Erica V. P. Lewis, Kelly Salchow Macarthur, Steven McCarthy, Shelly Mayers, Elizabeth Shefrin, Angelica Sibrian, David Smart, Ane Thon Knutsen, Isobel Thomas, Darryl Westley. - Coping with Covid: Pedagogy, teaching and learning: Bernard J Canniffe, Subir Dey, Aaron Ganci, Elizabeth Herrmann, John Kilburn, Paul Nini, Emily Osborne, Gianni Sinni & Irene Sgarro, Dave Wood, Helena Gregory, Colin Raeburn & Jackie Malcolm. - Coping with Covid: Social justice, activism and doing good: Class Action Collective, Xinyi Li, Matt Soar, Junie Tang, Lisa Winstanley. - Coping with Covid: Society, control and conspiracy: Diana Bîrhală, Maria Borțoi, Patti Capaldi, Tânia A. Cardoso, Peter Gibbons, Bianca Milea, Rebecca Tegtmeyer, Danne Wo

Hiv-1 Adapts To A Retrocyclin With Cationic Amino Acid Substitutions That Reduce Fusion Efficiency Of Gp41

Retrocyclin (RC)-101 is a cationic θ-defensin that inhibits HIV-1 entry. Passaging HIV-1BAL under selective pressure by this cyclic minidefensin resulted in only a 5- to 10-fold decrease in viral susceptibility to RC-101. Emergent viral isolates had three amino acid substitutions in their envelope glycoprotein. One was in a CD4-binding region of gp120, and the others were in the heptad repeat (HR) domains of gp41 (HR1 and HR2). Each mutation replaced an electroneutral or electronegative residue with one that was positively charged. These mutations were evaluated either alone or in combination in a single-round viral entry assay. Although the mutation in gp120 did not affect viral entry, the mutation in HR1 of gp41 conferred relative resistance to RC-101. Interestingly, the envelope with the HR2 mutation was less efficient and became codependent on the presence of RC-101 for entry. The adaptive response of HIV-1 to this cationic host defense peptide resembles the responses of bacteria that modulate their surface or membrane charge to evade analogous host defense peptides. These findings also suggest that interactions between θ-defensins and gp41 may contribute to the ability of these cyclic minidefensins to prevent HIV-1 entry into target cells. Copyright © 2006 by The American Association of Immunologists, Inc